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  • 99
    Millipore anti pfs25 4b7 mabs
    Anti Pfs25 4b7 Mabs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BEI Resources monoclonal anti pf s25 antibody 4b7
    Monoclonal Anti Pf S25 Antibody 4b7, supplied by BEI Resources, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Lampire Biological pf s25 based recombinant proteins
    Pf MSP8 is expressed during P. falciparum gametocyte development but is absent on the surface of activated macrogametes. (A) Detection of Pf MSP8 expression in fixed and permeabilized P. falciparum gametocytes (stages II to V) by an immunofluorescence assay with rabbit anti-r Pf MSP8 IgG or control IgG followed by FITC-conjugated secondary antibodies. DAPI was used to stain parasite DNA. (B) Analysis of Pf MSP8 expression on activated, live P. falciparum macrogametes by an immunofluorescence assay, as described above, with rabbit anti-r Pf MSP8 IgG. Samples were costained with MAb 4B7, which is specific for Pf <t>s25</t> (MAb 4B7), followed by TRITC-conjugated secondary IgG. DIC, differential interference contrast.
    Pf S25 Based Recombinant Proteins, supplied by Lampire Biological, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pfs25  (Kymab)
    91
    Kymab pfs25
    Molecular details of site 2 antibody recognition. a Superposition of site 2 <t>antibody–Pfs25</t> co-complex crystal structures. Pfs25s are shown as cartoon and colored according to EGF-like domain as in Fig. 3 . 1245 and 1260 Fabs are shown as surface representation and colored in salmon and brown, respectively. b Comparison of the angle of approach. The center of mass for both the Fab variable domains and interacting Pfs25 residues are shown as spheres and colored as in a . CDR loops that interact with Pfs25 for c 1245 and d 1260. Non-interacting Pfs25 residues are colored gray, while interacting residues are colored according to their EGF-like domain
    Pfs25, supplied by Kymab, used in various techniques. Bioz Stars score: 91/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    BEI Resources pfs25
    Changes in protein expression during formation and activation of P. falciparum gametocytes. ( A ) Immunofluorescence assays, using specific antibodies, detected the proteins of interest (in green) in asexual blood stage (ABS) parasites, in gametocytes (GC) and in gametocytes at 30 min p.a. (aGC). The parasite stages were highlighted with antibodies against stage-specific markers (in red; MSP1 for ABS; Pfs230 or <t>Pfs25</t> for GCs and aGCs). Nuclei were highlighted by Hoechst nuclear stain (in blue). Bar, 5 μm. ( B ) Diagram showing the average signal intensity of the immunolabeled proteins in gametocytes before and at 30 min p.a. in vitro. Measurements were performed on 20 plotted cells per setting via quantitative confocal microscopy. Mean ± SD. *p
    Pfs25, supplied by BEI Resources, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Novartis pfs25
    Changes in protein expression during formation and activation of P. falciparum gametocytes. ( A ) Immunofluorescence assays, using specific antibodies, detected the proteins of interest (in green) in asexual blood stage (ABS) parasites, in gametocytes (GC) and in gametocytes at 30 min p.a. (aGC). The parasite stages were highlighted with antibodies against stage-specific markers (in red; MSP1 for ABS; Pfs230 or <t>Pfs25</t> for GCs and aGCs). Nuclei were highlighted by Hoechst nuclear stain (in blue). Bar, 5 μm. ( B ) Diagram showing the average signal intensity of the immunolabeled proteins in gametocytes before and at 30 min p.a. in vitro. Measurements were performed on 20 plotted cells per setting via quantitative confocal microscopy. Mean ± SD. *p
    Pfs25, supplied by Novartis, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources p falciparum surface protein pfs25
    Changes in protein expression during formation and activation of P. falciparum gametocytes. ( A ) Immunofluorescence assays, using specific antibodies, detected the proteins of interest (in green) in asexual blood stage (ABS) parasites, in gametocytes (GC) and in gametocytes at 30 min p.a. (aGC). The parasite stages were highlighted with antibodies against stage-specific markers (in red; MSP1 for ABS; Pfs230 or <t>Pfs25</t> for GCs and aGCs). Nuclei were highlighted by Hoechst nuclear stain (in blue). Bar, 5 μm. ( B ) Diagram showing the average signal intensity of the immunolabeled proteins in gametocytes before and at 30 min p.a. in vitro. Measurements were performed on 20 plotted cells per setting via quantitative confocal microscopy. Mean ± SD. *p
    P Falciparum Surface Protein Pfs25, supplied by BEI Resources, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Fraunhofer USA pfs25 based immunogens
    Intensity of Plasmodium falciparum oocyst infections in a representative SMFA. The results are shown for one of 74 SMFAs conducted for this study. Each point represents the number of oocysts per mosquito and the black bar indicates the geometric mean for each group. Boxes are 95% confidence intervals around the geometric means. C is non-immune control serum (O + human serum). NC is negative control serum (pooled pre-immune mouse serum). S1 to S6 are serum samples from animals immunized with <t>Pfs25</t> immunogens.
    Pfs25 Based Immunogens, supplied by Fraunhofer USA, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millipore recombinant pfs25
    Mucosal antibody responses induced by intranasal immunization of mice with <t>Pfs25/CT,</t> CT alone, or PBS were analyzed by ELISA. Nasal wash samples were collected immediately after exsanguination by washing the nasal cavities several times with 200 μl
    Recombinant Pfs25, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    GenScript pfs25 gene
    <t>Pfs25-specific</t> serum antibody response upon heterologous prime-boost vaccination. a Vaccination schematic. Mice (n = 6 per group) were immunized with yPfs25-alum (2.5 μg/mouse) or Ad5 vector Pfs25 (10 9 particles) on day 0, and followed by boost immunization with yPfs25-alum (2.5 μg/mouse) or the different Ad5-HVR- pfs25 vectors (10 10 particles). The serum samples were collected 21 days after the boost. b yPfs25-specific antibody titer. yPfs25-specific ELISA was performed and data depict pooled serum from 6 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. c yPfs25-specific antibody avidity. The assay was performed as described for Fig. 3 . The fraction of Ig bound was determined by dividing the OD at each NaSCN concentration by the OD in the absence of NaSCN and the nonlinear best-fit line was plotted for each data set. d , e yPfs25-specific IgG sublass profile. The assay was performed as described for Fig. 3 . The average OD and standard deviation for each isotype/subclass ( d ) and the ratio of the O.D. for the IgG1 to the O.D.s of IgG2b and IgG3 ( e ) are shown. Data depict the ratio of the OD for IgG1 to the ODs of IgG2b and IgG3
    Pfs25 Gene, supplied by GenScript, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Fraunhofer USA pfs25 vlp fhcmb vaccine
    <t>Pfs25-specific</t> serum antibody response upon heterologous prime-boost vaccination. a Vaccination schematic. Mice (n = 6 per group) were immunized with yPfs25-alum (2.5 μg/mouse) or Ad5 vector Pfs25 (10 9 particles) on day 0, and followed by boost immunization with yPfs25-alum (2.5 μg/mouse) or the different Ad5-HVR- pfs25 vectors (10 10 particles). The serum samples were collected 21 days after the boost. b yPfs25-specific antibody titer. yPfs25-specific ELISA was performed and data depict pooled serum from 6 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. c yPfs25-specific antibody avidity. The assay was performed as described for Fig. 3 . The fraction of Ig bound was determined by dividing the OD at each NaSCN concentration by the OD in the absence of NaSCN and the nonlinear best-fit line was plotted for each data set. d , e yPfs25-specific IgG sublass profile. The assay was performed as described for Fig. 3 . The average OD and standard deviation for each isotype/subclass ( d ) and the ratio of the O.D. for the IgG1 to the O.D.s of IgG2b and IgG3 ( e ) are shown. Data depict the ratio of the OD for IgG1 to the ODs of IgG2b and IgG3
    Pfs25 Vlp Fhcmb Vaccine, supplied by Fraunhofer USA, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Bio-Rad pfs25 gene transcripts
    Plasmodium falciparum <t>Pfs25</t> detection from RDT by direct real time-PCR. a DNA amplification kinetics per cycle on plate 1 for each sample. b logarithmic value of fluorescence and presence of background noise. c Amplification on plate 2, d DNA amplification at 8 cycles. Reactions performed on a conventional RT-PCR instrument (Bio-Rad CFX Connect) with a threshold setting of 50 relative fluorescence units (horizontal line)
    Pfs25 Gene Transcripts, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher pfs25 cp sequence
    Schematic diagram of the ‘launch’ vector. Following agroinfiltration of plants, the sequence between the left border (LB) and the right border (RB) of the plasmid vector is transferred from Agrobacteria into plant cells where expression of the engineered TMV genome is driven by the Cauliflower mosaic virus (CaMV) 35S promoter. TMV replicase then drives amplification of primary transcript, and <t>Pfs25-CP</t> accumulation is then driven by the TMV CP subgenomic promoter (light blue box). Movement protein (MP) facilitates cell-to-cell transfer of viral sequences and is driven by the MP subgenomic promoter (dark blue box).
    Pfs25 Cp Sequence, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BEI Resources rabbit anti pfs25
    The effect of TSA on P. falciparum sexual stage development. (A) The effect of TSA on gametocyte development. TSA at IC 50 or IC 90 concentrations was added to stage II gametocyte cultures for 2 days. The numbers of stage IV and V gametocytes were determined in 1,000 erythrocytes at day 10 using Giemsa-stained blood smears. Epoxomicin (60 nM) was used as a positive control (not shown), while 0.5% vol. ethanol and chloroquine (16 nM) were used as negative controls (ethanol set to 100%). (B) The effect of TSA on macrogamete development. A mature gametocyte culture was incubated with TSA at IC 50 or IC 90 concentrations or 0.5% vol. ethanol for 1 h at 37°C. The culture was then activated with 100 μM XA and further cultured for 30 min at RT for macrogamete development. Macrogametes were detected by immunolabelling with <t>anti-Pfs25</t> and counted in triplicate in 1,000 erythrocytes. (C) Effect of TSA on zygote development. The zygote development assay was performed in the same way as the macrogamete development except for the fact that after activation the cultures were incubated for 12 h at RT. Results shown (for A–C ) are combined from three independent experiments each performed in triplicate (mean ± SD). * P
    Rabbit Anti Pfs25, supplied by BEI Resources, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources anti pfs25 igg
    The effect of TSA on P. falciparum sexual stage development. (A) The effect of TSA on gametocyte development. TSA at IC 50 or IC 90 concentrations was added to stage II gametocyte cultures for 2 days. The numbers of stage IV and V gametocytes were determined in 1,000 erythrocytes at day 10 using Giemsa-stained blood smears. Epoxomicin (60 nM) was used as a positive control (not shown), while 0.5% vol. ethanol and chloroquine (16 nM) were used as negative controls (ethanol set to 100%). (B) The effect of TSA on macrogamete development. A mature gametocyte culture was incubated with TSA at IC 50 or IC 90 concentrations or 0.5% vol. ethanol for 1 h at 37°C. The culture was then activated with 100 μM XA and further cultured for 30 min at RT for macrogamete development. Macrogametes were detected by immunolabelling with <t>anti-Pfs25</t> and counted in triplicate in 1,000 erythrocytes. (C) Effect of TSA on zygote development. The zygote development assay was performed in the same way as the macrogamete development except for the fact that after activation the cultures were incubated for 12 h at RT. Results shown (for A–C ) are combined from three independent experiments each performed in triplicate (mean ± SD). * P
    Anti Pfs25 Igg, supplied by BEI Resources, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    BEI Resources anti pfs25 mab 4b7
    SDS-PAGE and Western blot <t>(4B7)</t> of Pichia and baculovirus <t>Pfs25.</t> a SDS-PAGE and b Western blot using 4B7 monoclonal antibody of purified Pfs25 from Pichia under reducing conditions (non-reduced not show). c SDS-PAGE and d Western blot using 4B7 monoclonal antibody of non-reduced and reduced purified Pfs25 from baculovirus
    Anti Pfs25 Mab 4b7, supplied by BEI Resources, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Promega pfs25 required rq1 dnase i treatment
    SDS-PAGE and Western blot <t>(4B7)</t> of Pichia and baculovirus <t>Pfs25.</t> a SDS-PAGE and b Western blot using 4B7 monoclonal antibody of purified Pfs25 from Pichia under reducing conditions (non-reduced not show). c SDS-PAGE and d Western blot using 4B7 monoclonal antibody of non-reduced and reduced purified Pfs25 from baculovirus
    Pfs25 Required Rq1 Dnase I Treatment, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BioReliance grade pfs25 epa alhydrogel vaccine
    Characterization of <t>Pfs25-EPA</t> conjugate desorbed from <t>Alhydrogel</t> formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.
    Grade Pfs25 Epa Alhydrogel Vaccine, supplied by BioReliance, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    BEI Resources monoclonal anti pfs25 antibody38 4b7
    Characterization of <t>Pfs25-EPA</t> conjugate desorbed from <t>Alhydrogel</t> formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.
    Monoclonal Anti Pfs25 Antibody38 4b7, supplied by BEI Resources, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    ImmunoReagents polyclonal mouse anti pfs25 antibodies
    Characterization of <t>Pfs25-EPA</t> conjugate desorbed from <t>Alhydrogel</t> formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.
    Polyclonal Mouse Anti Pfs25 Antibodies, supplied by ImmunoReagents, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Mine Safety Appliances mouse serum albumin pfs25 msa
    Characterization of <t>Pfs25-EPA</t> conjugate desorbed from <t>Alhydrogel</t> formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.
    Mouse Serum Albumin Pfs25 Msa, supplied by Mine Safety Appliances, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore escherichia coli pfs25
    Characterization of <t>Pfs25-EPA</t> conjugate desorbed from <t>Alhydrogel</t> formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.
    Escherichia Coli Pfs25, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Pf MSP8 is expressed during P. falciparum gametocyte development but is absent on the surface of activated macrogametes. (A) Detection of Pf MSP8 expression in fixed and permeabilized P. falciparum gametocytes (stages II to V) by an immunofluorescence assay with rabbit anti-r Pf MSP8 IgG or control IgG followed by FITC-conjugated secondary antibodies. DAPI was used to stain parasite DNA. (B) Analysis of Pf MSP8 expression on activated, live P. falciparum macrogametes by an immunofluorescence assay, as described above, with rabbit anti-r Pf MSP8 IgG. Samples were costained with MAb 4B7, which is specific for Pf s25 (MAb 4B7), followed by TRITC-conjugated secondary IgG. DIC, differential interference contrast.

    Journal: Infection and Immunity

    Article Title: Evaluation of a Plasmodium-Specific Carrier Protein To Enhance Production of Recombinant Pfs25, a Leading Transmission-Blocking Vaccine Candidate

    doi: 10.1128/IAI.00486-17

    Figure Lengend Snippet: Pf MSP8 is expressed during P. falciparum gametocyte development but is absent on the surface of activated macrogametes. (A) Detection of Pf MSP8 expression in fixed and permeabilized P. falciparum gametocytes (stages II to V) by an immunofluorescence assay with rabbit anti-r Pf MSP8 IgG or control IgG followed by FITC-conjugated secondary antibodies. DAPI was used to stain parasite DNA. (B) Analysis of Pf MSP8 expression on activated, live P. falciparum macrogametes by an immunofluorescence assay, as described above, with rabbit anti-r Pf MSP8 IgG. Samples were costained with MAb 4B7, which is specific for Pf s25 (MAb 4B7), followed by TRITC-conjugated secondary IgG. DIC, differential interference contrast.

    Article Snippet: Polyclonal antisera were raised against Pf s25-based recombinant proteins by Lampire Biological Laboratories (Pipersville, PA).

    Techniques: Expressing, Immunofluorescence, Staining

    Purification of the r Pf s25-based vaccine. (A and B) Purified r Pf s25 was analyzed by SDS-PAGE (12% gel) under both reducing (R) and nonreducing (NR) conditions, followed by Coomassie blue staining (3 μg/lane) (A) or immunoblot analysis (0.2 μg/lane) (B) using anti- Pf s25 MAb 4B7. (C to E) Purified r Pfs 25/8(CΔS) was analyzed by SDS-PAGE (10% gel) under both reducing and nonreducing conditions, followed by Coomassie blue staining (3 μg/lane) (C) or immunoblot analysis (0.4 μg/lane) using anti- Pf s25 MAb 4B7 (D) or rabbit anti- Pf MSP8 (E).

    Journal: Infection and Immunity

    Article Title: Evaluation of a Plasmodium-Specific Carrier Protein To Enhance Production of Recombinant Pfs25, a Leading Transmission-Blocking Vaccine Candidate

    doi: 10.1128/IAI.00486-17

    Figure Lengend Snippet: Purification of the r Pf s25-based vaccine. (A and B) Purified r Pf s25 was analyzed by SDS-PAGE (12% gel) under both reducing (R) and nonreducing (NR) conditions, followed by Coomassie blue staining (3 μg/lane) (A) or immunoblot analysis (0.2 μg/lane) (B) using anti- Pf s25 MAb 4B7. (C to E) Purified r Pfs 25/8(CΔS) was analyzed by SDS-PAGE (10% gel) under both reducing and nonreducing conditions, followed by Coomassie blue staining (3 μg/lane) (C) or immunoblot analysis (0.4 μg/lane) using anti- Pf s25 MAb 4B7 (D) or rabbit anti- Pf MSP8 (E).

    Article Snippet: Polyclonal antisera were raised against Pf s25-based recombinant proteins by Lampire Biological Laboratories (Pipersville, PA).

    Techniques: Purification, SDS Page, Staining

    Immunization with r Pf s25 or r Pf s25/8(CΔS) elicits high titers of Pf s25-specific antibodies. New Zealand White rabbits were immunized three times with r Pf s25 ( n = 4) (black bars), r Pf s25/8(CΔS) ( n = 4) (white bars), antigens formulated with Alhydrogel as an adjuvant, or the adjuvant alone. Sera collected 2 weeks following each immunization were analyzed for antigen-specific IgG titers (means ± standard deviations) by an ELISA using plates coated with r Pf s25, r Pf s25/8(CΔS), or r Pf MSP8 antigens. The signal using adjuvant control sera was subtracted as the background. ns, nonsignificant ( P > 0.05).

    Journal: Infection and Immunity

    Article Title: Evaluation of a Plasmodium-Specific Carrier Protein To Enhance Production of Recombinant Pfs25, a Leading Transmission-Blocking Vaccine Candidate

    doi: 10.1128/IAI.00486-17

    Figure Lengend Snippet: Immunization with r Pf s25 or r Pf s25/8(CΔS) elicits high titers of Pf s25-specific antibodies. New Zealand White rabbits were immunized three times with r Pf s25 ( n = 4) (black bars), r Pf s25/8(CΔS) ( n = 4) (white bars), antigens formulated with Alhydrogel as an adjuvant, or the adjuvant alone. Sera collected 2 weeks following each immunization were analyzed for antigen-specific IgG titers (means ± standard deviations) by an ELISA using plates coated with r Pf s25, r Pf s25/8(CΔS), or r Pf MSP8 antigens. The signal using adjuvant control sera was subtracted as the background. ns, nonsignificant ( P > 0.05).

    Article Snippet: Polyclonal antisera were raised against Pf s25-based recombinant proteins by Lampire Biological Laboratories (Pipersville, PA).

    Techniques: Enzyme-linked Immunosorbent Assay

    Expression of chimeric r Pf s25/8(CΔS) and unfused r Pf s25. (A) Schematic depiction of expression constructs for the production of r Pf s25/8(CΔS) and r Pf s25. Expression plasmids for chimeric r Pf s25/8(CΔS) or unfused r Pf s25 were transformed into E. coli SHuffle T7 Express lysY cells. (B and C) r Pf s25/8(CΔS) lysates harvested before ( T 0 ) or 3 h after ( T 3 ) induction were separated by SDS-PAGE (10% gel) under reducing conditions, followed by Coomassie blue staining (B) or immunoblot analysis (C) using rabbit-anti-r Pf MSP8 IgG. The asterisk highlights r Pf s25/8(CΔS) at the predicted size. (D and E) The expression of unfused r Pf s25 was assessed as described above, on 12% polyacrylamide gels under reducing conditions, followed by Coomassie blue staining (D) or immunoblot analysis (E) using an anti-His MAb. The asterisk highlights r Pf s25 at the predicted size.

    Journal: Infection and Immunity

    Article Title: Evaluation of a Plasmodium-Specific Carrier Protein To Enhance Production of Recombinant Pfs25, a Leading Transmission-Blocking Vaccine Candidate

    doi: 10.1128/IAI.00486-17

    Figure Lengend Snippet: Expression of chimeric r Pf s25/8(CΔS) and unfused r Pf s25. (A) Schematic depiction of expression constructs for the production of r Pf s25/8(CΔS) and r Pf s25. Expression plasmids for chimeric r Pf s25/8(CΔS) or unfused r Pf s25 were transformed into E. coli SHuffle T7 Express lysY cells. (B and C) r Pf s25/8(CΔS) lysates harvested before ( T 0 ) or 3 h after ( T 3 ) induction were separated by SDS-PAGE (10% gel) under reducing conditions, followed by Coomassie blue staining (B) or immunoblot analysis (C) using rabbit-anti-r Pf MSP8 IgG. The asterisk highlights r Pf s25/8(CΔS) at the predicted size. (D and E) The expression of unfused r Pf s25 was assessed as described above, on 12% polyacrylamide gels under reducing conditions, followed by Coomassie blue staining (D) or immunoblot analysis (E) using an anti-His MAb. The asterisk highlights r Pf s25 at the predicted size.

    Article Snippet: Polyclonal antisera were raised against Pf s25-based recombinant proteins by Lampire Biological Laboratories (Pipersville, PA).

    Techniques: Expressing, Construct, Transformation Assay, SDS Page, Staining

    Molecular details of site 2 antibody recognition. a Superposition of site 2 antibody–Pfs25 co-complex crystal structures. Pfs25s are shown as cartoon and colored according to EGF-like domain as in Fig. 3 . 1245 and 1260 Fabs are shown as surface representation and colored in salmon and brown, respectively. b Comparison of the angle of approach. The center of mass for both the Fab variable domains and interacting Pfs25 residues are shown as spheres and colored as in a . CDR loops that interact with Pfs25 for c 1245 and d 1260. Non-interacting Pfs25 residues are colored gray, while interacting residues are colored according to their EGF-like domain

    Journal: Nature Communications

    Article Title: Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25

    doi: 10.1038/s41467-017-01924-3

    Figure Lengend Snippet: Molecular details of site 2 antibody recognition. a Superposition of site 2 antibody–Pfs25 co-complex crystal structures. Pfs25s are shown as cartoon and colored according to EGF-like domain as in Fig. 3 . 1245 and 1260 Fabs are shown as surface representation and colored in salmon and brown, respectively. b Comparison of the angle of approach. The center of mass for both the Fab variable domains and interacting Pfs25 residues are shown as spheres and colored as in a . CDR loops that interact with Pfs25 for c 1245 and d 1260. Non-interacting Pfs25 residues are colored gray, while interacting residues are colored according to their EGF-like domain

    Article Snippet: Two hundred twenty-five mAbs were confirmed by both assays to bind Pfs25, and had affinities ranging from ~500 nM to less than 1 nM (Supplementary Fig. ).

    Techniques:

    Affinity maturation of mAbs at the binding interface. Somatic hypermutations (SHMs) that introduce new contacts with Pfs25 are colored dark gray. a A S31R mutation in the HCDR1 of 1269 introduces two H-bonds with Pfs25 residues T93 and Q125. b A G55D mutation in the HCDR2 of 1269 introduces an additional H-bond with Pfs25 residue N87. c A S30N mutation in the LCDR1 of 1262 leads to an additional H-bond with Pfs25 residue N105. d A S54D mutation in the HCDR2 of 1190 leads to an additional H-bond contact with Pfs25 residue Q123. e A S52Y mutation in the LCDR2 of 1190 leads to additional van der Waals interactions with Pfs25 residues, and ensures that N87 hydrogen bonds with adjacent LCDR2 residues. f Similar to 1190, 1276 has a S52F mutation in the LCDR2 that leads to additional van der Waals contacts with Pfs25 residues, and ensures that N87 hydrogen bonds with adjacent LCDR2 residues

    Journal: Nature Communications

    Article Title: Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25

    doi: 10.1038/s41467-017-01924-3

    Figure Lengend Snippet: Affinity maturation of mAbs at the binding interface. Somatic hypermutations (SHMs) that introduce new contacts with Pfs25 are colored dark gray. a A S31R mutation in the HCDR1 of 1269 introduces two H-bonds with Pfs25 residues T93 and Q125. b A G55D mutation in the HCDR2 of 1269 introduces an additional H-bond with Pfs25 residue N87. c A S30N mutation in the LCDR1 of 1262 leads to an additional H-bond with Pfs25 residue N105. d A S54D mutation in the HCDR2 of 1190 leads to an additional H-bond contact with Pfs25 residue Q123. e A S52Y mutation in the LCDR2 of 1190 leads to additional van der Waals interactions with Pfs25 residues, and ensures that N87 hydrogen bonds with adjacent LCDR2 residues. f Similar to 1190, 1276 has a S52F mutation in the LCDR2 that leads to additional van der Waals contacts with Pfs25 residues, and ensures that N87 hydrogen bonds with adjacent LCDR2 residues

    Article Snippet: Two hundred twenty-five mAbs were confirmed by both assays to bind Pfs25, and had affinities ranging from ~500 nM to less than 1 nM (Supplementary Fig. ).

    Techniques: Binding Assay, Introduce, Mutagenesis

    Phylogram representation of B-cell responses to Pfs25-VLP immunization. Natively paired H- and L-chain, full-length, variable region sequences from 13 Pfs25-VLP-immunized Kymice were combined into one phylogram based on full-variable region H- and L-chain amino acid sequence similarity among plasmablast IgG and memory B-cell IgG sequences. The 95 plasmablast-derived and 18 memory B-cell-derived sequences selected for recombinant antibody expression are noted as green circles and blue boxes, respectively. Recombinant antibodies listed in Table 1 chosen for further analysis are labeled. Between pairs of nodes, sum of radial distances is proportional to the Kimura distance as indicated by the scale bar. Branch color denotes the specific animal from which the sequence was derived. Higher magnification insets show cases in which two different anti-Pfs25 antibodies of highly similar sequence originate from different individual Kymice and, in two cases, different antibody-expressing cell types. The subset of plasmablast sequences from Pfs25-VLP immunized Kymice that are similar to sequences from empty VLP-immunized Kymice are included as part of the 1564 plasmablast data set shown in the tree, but were not used as sources for recombinant antibody expression

    Journal: Nature Communications

    Article Title: Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25

    doi: 10.1038/s41467-017-01924-3

    Figure Lengend Snippet: Phylogram representation of B-cell responses to Pfs25-VLP immunization. Natively paired H- and L-chain, full-length, variable region sequences from 13 Pfs25-VLP-immunized Kymice were combined into one phylogram based on full-variable region H- and L-chain amino acid sequence similarity among plasmablast IgG and memory B-cell IgG sequences. The 95 plasmablast-derived and 18 memory B-cell-derived sequences selected for recombinant antibody expression are noted as green circles and blue boxes, respectively. Recombinant antibodies listed in Table 1 chosen for further analysis are labeled. Between pairs of nodes, sum of radial distances is proportional to the Kimura distance as indicated by the scale bar. Branch color denotes the specific animal from which the sequence was derived. Higher magnification insets show cases in which two different anti-Pfs25 antibodies of highly similar sequence originate from different individual Kymice and, in two cases, different antibody-expressing cell types. The subset of plasmablast sequences from Pfs25-VLP immunized Kymice that are similar to sequences from empty VLP-immunized Kymice are included as part of the 1564 plasmablast data set shown in the tree, but were not used as sources for recombinant antibody expression

    Article Snippet: Two hundred twenty-five mAbs were confirmed by both assays to bind Pfs25, and had affinities ranging from ~500 nM to less than 1 nM (Supplementary Fig. ).

    Techniques: Sequencing, Derivative Assay, Recombinant, Expressing, Labeling

    Epitope binning of anti-Pfs25 Kymouse-derived human antibodies. Primary antibodies tested are listed in the left column, while secondary competing antibodies are listed at the top in rows. Data indicate the percent of competing antibody binding compared to the maximum competing antibody response in the absence of the primary antibody. Boxes are colored according to competition status. Competing antibodies that displayed ≤ 33% maximal binding are colored black and considered competing, between 34 and 66% binding are considered intermediate and colored gray, and those that displayed ≥ 66% binding are colored white and considered non-competing. Epitope bins are listed above or beside the table, with site 1 competing antibodies colored in shades of blue and site 2 competing antibodies colored green. *Primary antibodies displayed a fast off-rate, resulting in intermediate binding values for true non-competing antibodies. ^Competing antibodies displayed a slow on-rate, leading to low values. The well-characterized mouse mAb 4B7 is highlighted in yellow as a reference

    Journal: Nature Communications

    Article Title: Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25

    doi: 10.1038/s41467-017-01924-3

    Figure Lengend Snippet: Epitope binning of anti-Pfs25 Kymouse-derived human antibodies. Primary antibodies tested are listed in the left column, while secondary competing antibodies are listed at the top in rows. Data indicate the percent of competing antibody binding compared to the maximum competing antibody response in the absence of the primary antibody. Boxes are colored according to competition status. Competing antibodies that displayed ≤ 33% maximal binding are colored black and considered competing, between 34 and 66% binding are considered intermediate and colored gray, and those that displayed ≥ 66% binding are colored white and considered non-competing. Epitope bins are listed above or beside the table, with site 1 competing antibodies colored in shades of blue and site 2 competing antibodies colored green. *Primary antibodies displayed a fast off-rate, resulting in intermediate binding values for true non-competing antibodies. ^Competing antibodies displayed a slow on-rate, leading to low values. The well-characterized mouse mAb 4B7 is highlighted in yellow as a reference

    Article Snippet: Two hundred twenty-five mAbs were confirmed by both assays to bind Pfs25, and had affinities ranging from ~500 nM to less than 1 nM (Supplementary Fig. ).

    Techniques: Derivative Assay, Binding Assay

    Molecular details of site 1a antibody recognition. a Superposition of site 1a antibody–Pfs25 co-complex crystal structures. Pfs25’s are shown as cartoon and colored according to EGF-like domain as in Fig. 3 . 1190, 1262, 1269, and 1276 Fabs are shown as surface representation and colored in yellow, blue, orange, and teal, respectively. b Comparison of the angle of approach. The center of mass for both the Fab variable domains and interacting Pfs25 residues are shown as spheres and colored as in a . CDR loops that interact with Pfs25 for c 1190, d 1276, e 1262, and f 1269. Non-interacting Pfs25 residues are colored gray, while interacting residues are colored according to their EGF-like domain as in a

    Journal: Nature Communications

    Article Title: Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25

    doi: 10.1038/s41467-017-01924-3

    Figure Lengend Snippet: Molecular details of site 1a antibody recognition. a Superposition of site 1a antibody–Pfs25 co-complex crystal structures. Pfs25’s are shown as cartoon and colored according to EGF-like domain as in Fig. 3 . 1190, 1262, 1269, and 1276 Fabs are shown as surface representation and colored in yellow, blue, orange, and teal, respectively. b Comparison of the angle of approach. The center of mass for both the Fab variable domains and interacting Pfs25 residues are shown as spheres and colored as in a . CDR loops that interact with Pfs25 for c 1190, d 1276, e 1262, and f 1269. Non-interacting Pfs25 residues are colored gray, while interacting residues are colored according to their EGF-like domain as in a

    Article Snippet: Two hundred twenty-five mAbs were confirmed by both assays to bind Pfs25, and had affinities ranging from ~500 nM to less than 1 nM (Supplementary Fig. ).

    Techniques:

    Crystal structures of six mAbs in complex with Pfs25. Crystal structures of a 1190, b 1276, c 1262, d 1269, e 1245, and f 1260 in complex with Pfs25. All crystal structures are shown according to the same Pfs25 orientation. Pfs25 is represented as surface and EGF-like domains 1–4 are colored in wheat, pink, green, and blue, respectively. g Site 1a epitopes for 1190, 1262, 1269, and 1276 antibodies traced onto the surface of Pfs25. h Site 2 epitopes for 1245 and 1260 antibodies traced onto the surface of Pfs25

    Journal: Nature Communications

    Article Title: Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25

    doi: 10.1038/s41467-017-01924-3

    Figure Lengend Snippet: Crystal structures of six mAbs in complex with Pfs25. Crystal structures of a 1190, b 1276, c 1262, d 1269, e 1245, and f 1260 in complex with Pfs25. All crystal structures are shown according to the same Pfs25 orientation. Pfs25 is represented as surface and EGF-like domains 1–4 are colored in wheat, pink, green, and blue, respectively. g Site 1a epitopes for 1190, 1262, 1269, and 1276 antibodies traced onto the surface of Pfs25. h Site 2 epitopes for 1245 and 1260 antibodies traced onto the surface of Pfs25

    Article Snippet: Two hundred twenty-five mAbs were confirmed by both assays to bind Pfs25, and had affinities ranging from ~500 nM to less than 1 nM (Supplementary Fig. ).

    Techniques:

    Changes in protein expression during formation and activation of P. falciparum gametocytes. ( A ) Immunofluorescence assays, using specific antibodies, detected the proteins of interest (in green) in asexual blood stage (ABS) parasites, in gametocytes (GC) and in gametocytes at 30 min p.a. (aGC). The parasite stages were highlighted with antibodies against stage-specific markers (in red; MSP1 for ABS; Pfs230 or Pfs25 for GCs and aGCs). Nuclei were highlighted by Hoechst nuclear stain (in blue). Bar, 5 μm. ( B ) Diagram showing the average signal intensity of the immunolabeled proteins in gametocytes before and at 30 min p.a. in vitro. Measurements were performed on 20 plotted cells per setting via quantitative confocal microscopy. Mean ± SD. *p

    Journal: BMC Genomics

    Article Title: Changes in the transcriptome of the malaria parasite Plasmodium falciparum during the initial phase of transmission from the human to the mosquito

    doi: 10.1186/1471-2164-14-256

    Figure Lengend Snippet: Changes in protein expression during formation and activation of P. falciparum gametocytes. ( A ) Immunofluorescence assays, using specific antibodies, detected the proteins of interest (in green) in asexual blood stage (ABS) parasites, in gametocytes (GC) and in gametocytes at 30 min p.a. (aGC). The parasite stages were highlighted with antibodies against stage-specific markers (in red; MSP1 for ABS; Pfs230 or Pfs25 for GCs and aGCs). Nuclei were highlighted by Hoechst nuclear stain (in blue). Bar, 5 μm. ( B ) Diagram showing the average signal intensity of the immunolabeled proteins in gametocytes before and at 30 min p.a. in vitro. Measurements were performed on 20 plotted cells per setting via quantitative confocal microscopy. Mean ± SD. *p

    Article Snippet: Antibodies The following antibodies were used in this study: mouse antisera against Pfs16 and Pfs230 (kindly provided by Kim Williamson, Loyola University Chicago), proteasome SU α5 [ ] and actin II [ ], as well as rabbit antisera against GAP50 (kindly provided by Julian Rayner, Sanger Institute England; [ ]), Pfs230 (against the immunogenic region C) [ ], MSP-1 and Pfs25 (BEI Resources, Manassas).

    Techniques: Expressing, Activation Assay, Immunofluorescence, Staining, Immunolabeling, In Vitro, Confocal Microscopy

    Immunogenicity of Pfs25-FhCMB in mice and rabbits

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Immunogenicity of Pfs25-FhCMB in mice and rabbits

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Mouse Assay

    Figure 1. ( A ) Schematic of the expression cassette for Pfs25-FhCMB. The PR-1a signal sequence is followed by the lichenase gene fused to the N-terminal of Pfs25. A 6xHis affinity purification tag and the KDEL sequence are cloned to the C-terminus

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Figure 1. ( A ) Schematic of the expression cassette for Pfs25-FhCMB. The PR-1a signal sequence is followed by the lichenase gene fused to the N-terminal of Pfs25. A 6xHis affinity purification tag and the KDEL sequence are cloned to the C-terminus

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Expressing, Sequencing, Affinity Purification, Clone Assay

    Immunogenicity of Pfs25-FhCMB in mice and rabbits

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Immunogenicity of Pfs25-FhCMB in mice and rabbits

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Mouse Assay

    Figure 5. Pfs25-FhCMB molecular weight determination by SEC-MALS and mass spectrometry. ( A ) Analytical SEC-MALS elution profile and ( B ) deconvoluted intact mass spectra for Pfs25-FhCMB showing a discrete, monomeric protein.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Figure 5. Pfs25-FhCMB molecular weight determination by SEC-MALS and mass spectrometry. ( A ) Analytical SEC-MALS elution profile and ( B ) deconvoluted intact mass spectra for Pfs25-FhCMB showing a discrete, monomeric protein.

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Molecular Weight, Size-exclusion Chromatography, Mass Spectrometry

    Figure 4. Electrophoretic analysis of purified Pfs25-FhCMB. Electrophoretic analysis included ( A ) reduced and ( B ) non-reduced Coomassie-stained SDS-PAGE and western blot analysis with ( C ) an anti-Pfs25 mAb 4B7, ( D ) an anti-LicKM polyclonal antiserum,

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Figure 4. Electrophoretic analysis of purified Pfs25-FhCMB. Electrophoretic analysis included ( A ) reduced and ( B ) non-reduced Coomassie-stained SDS-PAGE and western blot analysis with ( C ) an anti-Pfs25 mAb 4B7, ( D ) an anti-LicKM polyclonal antiserum,

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Purification, Staining, SDS Page, Western Blot

    Figure 2. Solubility and expression determination for Pfs25-FhCMB. Western blot analysis of total homogenate (TH), pellet suspended in 1x PBS, and total soluble protein (TSP) with ( A ) anti-4xHis mAb and ( B ) anti-LicKM polyclonal antiserum.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Figure 2. Solubility and expression determination for Pfs25-FhCMB. Western blot analysis of total homogenate (TH), pellet suspended in 1x PBS, and total soluble protein (TSP) with ( A ) anti-4xHis mAb and ( B ) anti-LicKM polyclonal antiserum.

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Solubility, Expressing, Western Blot

    Immunogenicity of Pfs25-FhCMB in mice and rabbits

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Immunogenicity of Pfs25-FhCMB in mice and rabbits

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Mouse Assay

    Figure 3. Pfs25-FhCMB purification schematic and in-process monitoring. ( A ) Process flow schematic for purification of Pfs25-FhCMB. ( B ) Reduced Coomassie-stained SDS-PAGE (10%) of purification process samples. M, molecular weight markers; H, homogenate;

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: A novel plant-produced Pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses

    doi: 10.4161/hv.34366

    Figure Lengend Snippet: Figure 3. Pfs25-FhCMB purification schematic and in-process monitoring. ( A ) Process flow schematic for purification of Pfs25-FhCMB. ( B ) Reduced Coomassie-stained SDS-PAGE (10%) of purification process samples. M, molecular weight markers; H, homogenate;

    Article Snippet: For western blot analysis, samples were transferred to polyvinylidene difluoride membranes, blocked with I-Block (Applied Biosystems), and developed using either anti-4xHis mAb (Qiagen), anti-LicKM polyclonal antiserum (produced by Fraunhofer Center for Molecular Biotechnology) or anti-Pfs25 mAb (4B7; obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Mus musculus (B cell); Mus musculus (myeloma) 4B7, MRA-315, deposited by LH Miller, A Saul).

    Techniques: Purification, Flow Cytometry, Staining, SDS Page, Molecular Weight

    Intensity of Plasmodium falciparum oocyst infections in a representative SMFA. The results are shown for one of 74 SMFAs conducted for this study. Each point represents the number of oocysts per mosquito and the black bar indicates the geometric mean for each group. Boxes are 95% confidence intervals around the geometric means. C is non-immune control serum (O + human serum). NC is negative control serum (pooled pre-immune mouse serum). S1 to S6 are serum samples from animals immunized with Pfs25 immunogens.

    Journal: Malaria Journal

    Article Title: Robust, reproducible, industrialized, standard membrane feeding assay for assessing the transmission blocking activity of vaccines and drugs against Plasmodium falciparum

    doi: 10.1186/s12936-015-0665-8

    Figure Lengend Snippet: Intensity of Plasmodium falciparum oocyst infections in a representative SMFA. The results are shown for one of 74 SMFAs conducted for this study. Each point represents the number of oocysts per mosquito and the black bar indicates the geometric mean for each group. Boxes are 95% confidence intervals around the geometric means. C is non-immune control serum (O + human serum). NC is negative control serum (pooled pre-immune mouse serum). S1 to S6 are serum samples from animals immunized with Pfs25 immunogens.

    Article Snippet: One-hundred and eighty-eight serum samples from mice immunized with Pfs25-based immunogens were obtained from Fraunhofer USA Center for Molecular Biotechnology (FhCMB) [ , ].

    Techniques: Negative Control

    Effect of blood meal quality on transmission blocking effect of two anti-Pfs25 monoclonal antibodies. The relationship between infection intensity (geometric mean number of oocysts) in control mosquitoes and the infection intensity (A, B) and infection prevalence (proportion of mosquitoes with oocysts) (C, D) is plotted for mosquitoes fed anti-Pfs25 antibodies, MRA39 (A, C) and MRA38 (B, D) in the infectious blood meals.

    Journal: Malaria Journal

    Article Title: Robust, reproducible, industrialized, standard membrane feeding assay for assessing the transmission blocking activity of vaccines and drugs against Plasmodium falciparum

    doi: 10.1186/s12936-015-0665-8

    Figure Lengend Snippet: Effect of blood meal quality on transmission blocking effect of two anti-Pfs25 monoclonal antibodies. The relationship between infection intensity (geometric mean number of oocysts) in control mosquitoes and the infection intensity (A, B) and infection prevalence (proportion of mosquitoes with oocysts) (C, D) is plotted for mosquitoes fed anti-Pfs25 antibodies, MRA39 (A, C) and MRA38 (B, D) in the infectious blood meals.

    Article Snippet: One-hundred and eighty-eight serum samples from mice immunized with Pfs25-based immunogens were obtained from Fraunhofer USA Center for Molecular Biotechnology (FhCMB) [ , ].

    Techniques: Transmission Assay, Blocking Assay, Infection

    Mucosal antibody responses induced by intranasal immunization of mice with Pfs25/CT, CT alone, or PBS were analyzed by ELISA. Nasal wash samples were collected immediately after exsanguination by washing the nasal cavities several times with 200 μl

    Journal:

    Article Title: Nasal Immunization with a Malaria Transmission-Blocking Vaccine Candidate, Pfs25, Induces Complete Protective Immunity in Mice against Field Isolates of Plasmodium falciparum

    doi: 10.1128/IAI.73.11.7375-7380.2005

    Figure Lengend Snippet: Mucosal antibody responses induced by intranasal immunization of mice with Pfs25/CT, CT alone, or PBS were analyzed by ELISA. Nasal wash samples were collected immediately after exsanguination by washing the nasal cavities several times with 200 μl

    Article Snippet: Groups of six to seven mice were intranasally immunized three times at weeks 0, 3, and 5 with 20 μg of Pichia pastoris -expressed recombinant Pfs25 ( ) mixed with 1 μg of cholera toxin (CT; Sigma-Aldrich) as a mucosal adjuvant.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Serum antibody responses induced by intranasal immunization of BALB/c and A/J mice with Pfs25/CT, CT alone, or PBS were analyzed by ELISA. Groups of six to seven mice were immunized three times at weeks 0, 3, and 5, and immune sera were collected at week

    Journal:

    Article Title: Nasal Immunization with a Malaria Transmission-Blocking Vaccine Candidate, Pfs25, Induces Complete Protective Immunity in Mice against Field Isolates of Plasmodium falciparum

    doi: 10.1128/IAI.73.11.7375-7380.2005

    Figure Lengend Snippet: Serum antibody responses induced by intranasal immunization of BALB/c and A/J mice with Pfs25/CT, CT alone, or PBS were analyzed by ELISA. Groups of six to seven mice were immunized three times at weeks 0, 3, and 5, and immune sera were collected at week

    Article Snippet: Groups of six to seven mice were intranasally immunized three times at weeks 0, 3, and 5 with 20 μg of Pichia pastoris -expressed recombinant Pfs25 ( ) mixed with 1 μg of cholera toxin (CT; Sigma-Aldrich) as a mucosal adjuvant.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Ookinete-specific reactivity of Pfs25/CT immune sera was confirmed by immunofluorescence analysis. The immune sera specifically recognized native Pfs25 proteins expressed on the surface of P. falciparum ookinetes. The immune sera did not react with gametocytes.

    Journal:

    Article Title: Nasal Immunization with a Malaria Transmission-Blocking Vaccine Candidate, Pfs25, Induces Complete Protective Immunity in Mice against Field Isolates of Plasmodium falciparum

    doi: 10.1128/IAI.73.11.7375-7380.2005

    Figure Lengend Snippet: Ookinete-specific reactivity of Pfs25/CT immune sera was confirmed by immunofluorescence analysis. The immune sera specifically recognized native Pfs25 proteins expressed on the surface of P. falciparum ookinetes. The immune sera did not react with gametocytes.

    Article Snippet: Groups of six to seven mice were intranasally immunized three times at weeks 0, 3, and 5 with 20 μg of Pichia pastoris -expressed recombinant Pfs25 ( ) mixed with 1 μg of cholera toxin (CT; Sigma-Aldrich) as a mucosal adjuvant.

    Techniques: Immunofluorescence

    Pfs25-specific serum antibody response upon heterologous prime-boost vaccination. a Vaccination schematic. Mice (n = 6 per group) were immunized with yPfs25-alum (2.5 μg/mouse) or Ad5 vector Pfs25 (10 9 particles) on day 0, and followed by boost immunization with yPfs25-alum (2.5 μg/mouse) or the different Ad5-HVR- pfs25 vectors (10 10 particles). The serum samples were collected 21 days after the boost. b yPfs25-specific antibody titer. yPfs25-specific ELISA was performed and data depict pooled serum from 6 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. c yPfs25-specific antibody avidity. The assay was performed as described for Fig. 3 . The fraction of Ig bound was determined by dividing the OD at each NaSCN concentration by the OD in the absence of NaSCN and the nonlinear best-fit line was plotted for each data set. d , e yPfs25-specific IgG sublass profile. The assay was performed as described for Fig. 3 . The average OD and standard deviation for each isotype/subclass ( d ) and the ratio of the O.D. for the IgG1 to the O.D.s of IgG2b and IgG3 ( e ) are shown. Data depict the ratio of the OD for IgG1 to the ODs of IgG2b and IgG3

    Journal: Malaria Journal

    Article Title: New adenovirus-based vaccine vectors targeting Pfs25 elicit antibodies that inhibit Plasmodium falciparum transmission

    doi: 10.1186/s12936-017-1896-7

    Figure Lengend Snippet: Pfs25-specific serum antibody response upon heterologous prime-boost vaccination. a Vaccination schematic. Mice (n = 6 per group) were immunized with yPfs25-alum (2.5 μg/mouse) or Ad5 vector Pfs25 (10 9 particles) on day 0, and followed by boost immunization with yPfs25-alum (2.5 μg/mouse) or the different Ad5-HVR- pfs25 vectors (10 10 particles). The serum samples were collected 21 days after the boost. b yPfs25-specific antibody titer. yPfs25-specific ELISA was performed and data depict pooled serum from 6 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. c yPfs25-specific antibody avidity. The assay was performed as described for Fig. 3 . The fraction of Ig bound was determined by dividing the OD at each NaSCN concentration by the OD in the absence of NaSCN and the nonlinear best-fit line was plotted for each data set. d , e yPfs25-specific IgG sublass profile. The assay was performed as described for Fig. 3 . The average OD and standard deviation for each isotype/subclass ( d ) and the ratio of the O.D. for the IgG1 to the O.D.s of IgG2b and IgG3 ( e ) are shown. Data depict the ratio of the OD for IgG1 to the ODs of IgG2b and IgG3

    Article Snippet: Briefly, the Pfs25 gene (bp 1–217) was codon-optimized for expression in humans (Genscript, Piscataway, NJ, USA) and inserted it into the pshuttle-CMV vector (Agilent Technologies plasmid #240007, Santa Clara, CA, USA).

    Techniques: Mouse Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Concentration Assay

    Vaccination induced transmission-blocking activity. Two independent sets of mice were immunized with the test groups, yPfs25 homologous prime boost (yPfs25-alum/yPfs25-alum) or the 3 heterologous prime boost strategies (Ad5- pfs25 /yPfs25-alum, Ad5- pfs25 /Ad5-HVR5D2, and Ad5- pfs25 /Ad5-HVR5D3) as described in

    Journal: Malaria Journal

    Article Title: New adenovirus-based vaccine vectors targeting Pfs25 elicit antibodies that inhibit Plasmodium falciparum transmission

    doi: 10.1186/s12936-017-1896-7

    Figure Lengend Snippet: Vaccination induced transmission-blocking activity. Two independent sets of mice were immunized with the test groups, yPfs25 homologous prime boost (yPfs25-alum/yPfs25-alum) or the 3 heterologous prime boost strategies (Ad5- pfs25 /yPfs25-alum, Ad5- pfs25 /Ad5-HVR5D2, and Ad5- pfs25 /Ad5-HVR5D3) as described in " Methods ". Serum was pooled from the test groups from each immunization (n = 3 mice for all the immunization groups except the 2nd yPfs25 homologous prime boost immunization, which included 6 mice) and tested in SMFAs at a 1:10 dilution. Pooled serum from mice immunized with an Ad5-luciferase vector was the negative control in each assay. The first SMFA tested pooled serum from the first immunization, while the second and third SMFAs tested pooled serum from the second immunization. All three SMFAs were included in the statistical analysis. The best estimate of the percent inhibition from 3 independent SMFAs is plotted with the 95% confidence interval for each sample calculated using a zero-inflated negative binomial model [ 30 ] (***p

    Article Snippet: Briefly, the Pfs25 gene (bp 1–217) was codon-optimized for expression in humans (Genscript, Piscataway, NJ, USA) and inserted it into the pshuttle-CMV vector (Agilent Technologies plasmid #240007, Santa Clara, CA, USA).

    Techniques: Transmission Assay, Blocking Assay, Activity Assay, Mouse Assay, Luciferase, Plasmid Preparation, Negative Control, Inhibition

    Pfs25 expression on Hela cells. Hela cells were transfected with an Ad5 shuttle vector with ( a ) or without ( b ) the codon-optimized Pfs25 gene under the control of the CMV immediate early promotor. Twenty-four hours post transfection, cells were fixed, permeabilized, and probed with ID2, a conformation-dependent Pfs25-specific monoclonal antibody, then probed with an Alexa Fluor488-conjugated anti-mouse secondary antibody and visualized by IFA

    Journal: Malaria Journal

    Article Title: New adenovirus-based vaccine vectors targeting Pfs25 elicit antibodies that inhibit Plasmodium falciparum transmission

    doi: 10.1186/s12936-017-1896-7

    Figure Lengend Snippet: Pfs25 expression on Hela cells. Hela cells were transfected with an Ad5 shuttle vector with ( a ) or without ( b ) the codon-optimized Pfs25 gene under the control of the CMV immediate early promotor. Twenty-four hours post transfection, cells were fixed, permeabilized, and probed with ID2, a conformation-dependent Pfs25-specific monoclonal antibody, then probed with an Alexa Fluor488-conjugated anti-mouse secondary antibody and visualized by IFA

    Article Snippet: Briefly, the Pfs25 gene (bp 1–217) was codon-optimized for expression in humans (Genscript, Piscataway, NJ, USA) and inserted it into the pshuttle-CMV vector (Agilent Technologies plasmid #240007, Santa Clara, CA, USA).

    Techniques: Expressing, Transfection, Plasmid Preparation, Immunofluorescence

    Pfs25 specific Ig production and T-cell activation following Ad5- pfs25 primary vaccination. Mice (n = 3 per group) were immunized with Ad5 expressing Pfs25 (10 10 particles) or recombinant yPfs25-alum (2.5 µg/mouse) on day 0, and serum and splenocyte samples were collected on day 10. a yPfs25-specific ELISA, yPfs25-specific ELISA were performed and data depict pooled serum from 3 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. b Splenocyte IFNγ secretion. Splenocytes (n = 3, tested individually) were incubated without or with yPfs25 at 10 μg/ml or Ad5 at 10,000 particles per cell for 24 h as indicated on the x - axis . IFNγ secretion was detected by ELISA and the average and standard deviation are plotted. Statistical significance was determined by Student’s T test; ***p

    Journal: Malaria Journal

    Article Title: New adenovirus-based vaccine vectors targeting Pfs25 elicit antibodies that inhibit Plasmodium falciparum transmission

    doi: 10.1186/s12936-017-1896-7

    Figure Lengend Snippet: Pfs25 specific Ig production and T-cell activation following Ad5- pfs25 primary vaccination. Mice (n = 3 per group) were immunized with Ad5 expressing Pfs25 (10 10 particles) or recombinant yPfs25-alum (2.5 µg/mouse) on day 0, and serum and splenocyte samples were collected on day 10. a yPfs25-specific ELISA, yPfs25-specific ELISA were performed and data depict pooled serum from 3 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. b Splenocyte IFNγ secretion. Splenocytes (n = 3, tested individually) were incubated without or with yPfs25 at 10 μg/ml or Ad5 at 10,000 particles per cell for 24 h as indicated on the x - axis . IFNγ secretion was detected by ELISA and the average and standard deviation are plotted. Statistical significance was determined by Student’s T test; ***p

    Article Snippet: Briefly, the Pfs25 gene (bp 1–217) was codon-optimized for expression in humans (Genscript, Piscataway, NJ, USA) and inserted it into the pshuttle-CMV vector (Agilent Technologies plasmid #240007, Santa Clara, CA, USA).

    Techniques: Activation Assay, Mouse Assay, Expressing, Recombinant, Enzyme-linked Immunosorbent Assay, Standard Deviation, Incubation

    Pfs25 epitope specific vectors design. a Amino acid sequence of Pfs25. The selected Pfs25 EGF domains II and III from which the epitope peptides (denoted as D2 and D3) were chosen for HVR insertion are indicated in green and orange , respectively. b Predicted Pfs25 structure based on the structure of the P. vivax homolog, Pvs25. The D2 and D3 epitopes in EGF domains II and III, respectively, are highlighted in red . c Ad5 Hexon capsid protein structure. Ad5 Hexon trimer structure (Protein Data Bank 1P30) with the modelled HVRs highlighted (HVR1, red ; HVR2, green ; HVR3, pink ; HVR4, light blue ; HVR5, yellow ; HVR6, blue ; HVR7, cyan ). d yPfs25-specific antibody titer. Mice (n = 3 per group) were immunized with 10 10 viral particles of Ad5 displaying the D2 or D3 Pfs25 peptide epitope within HVR5 (denoted Ad5-HVR5D2 or Ad5-HVR5D3, respectively) or PBS control and yPfs25-specific ELISA was performed and data depict endpoint dilutions for 3 individual animals where error bars depict standard deviation

    Journal: Malaria Journal

    Article Title: New adenovirus-based vaccine vectors targeting Pfs25 elicit antibodies that inhibit Plasmodium falciparum transmission

    doi: 10.1186/s12936-017-1896-7

    Figure Lengend Snippet: Pfs25 epitope specific vectors design. a Amino acid sequence of Pfs25. The selected Pfs25 EGF domains II and III from which the epitope peptides (denoted as D2 and D3) were chosen for HVR insertion are indicated in green and orange , respectively. b Predicted Pfs25 structure based on the structure of the P. vivax homolog, Pvs25. The D2 and D3 epitopes in EGF domains II and III, respectively, are highlighted in red . c Ad5 Hexon capsid protein structure. Ad5 Hexon trimer structure (Protein Data Bank 1P30) with the modelled HVRs highlighted (HVR1, red ; HVR2, green ; HVR3, pink ; HVR4, light blue ; HVR5, yellow ; HVR6, blue ; HVR7, cyan ). d yPfs25-specific antibody titer. Mice (n = 3 per group) were immunized with 10 10 viral particles of Ad5 displaying the D2 or D3 Pfs25 peptide epitope within HVR5 (denoted Ad5-HVR5D2 or Ad5-HVR5D3, respectively) or PBS control and yPfs25-specific ELISA was performed and data depict endpoint dilutions for 3 individual animals where error bars depict standard deviation

    Article Snippet: Briefly, the Pfs25 gene (bp 1–217) was codon-optimized for expression in humans (Genscript, Piscataway, NJ, USA) and inserted it into the pshuttle-CMV vector (Agilent Technologies plasmid #240007, Santa Clara, CA, USA).

    Techniques: Sequencing, Mouse Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Pfs25-specific serum antibody response after primary immunization with an Ad vector expressing Pfs25. Mice (n = 3 per group) were immunized with Ad5 expressing Pfs25 (10 10 particles) or recombinant yPfs25-alum (25 μg/mouse) on day 0, and serum samples were collected on day 21. a yPfs25-specific antibody titer. yPfs25-specific ELISA was performed and data depict pooled serum from 3 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. b yPfs25-specific avidity. Sera were added to a yPfs25-coated ELISA plate at a single dilution and then incubated with increasing concentrations of NaSCN. The remaining yPfs25-specific antibodies were detected with an HRP-conjugated anti-mouse IgG (Fc). The fraction of Ig bound was determined by dividing the optical density (OD) at each NaSCN concentration by the OD in the absence of NaSCN and the nonlinear best-fit line was plotted for each data set. c , d yPfs25-specific IgM and IgG subclass profile. yPfs25-specific ELISA was performed and the yPfs25-specific IgM and IgG subclasses were detected with HRP-conjugated specific antibodies. The average OD and standard deviation for each isotype/subclass ( c ) and the ratio of the O.D. for the IgG1 to the O.D.s of IgG2b and IgG3 ( d ) are shown

    Journal: Malaria Journal

    Article Title: New adenovirus-based vaccine vectors targeting Pfs25 elicit antibodies that inhibit Plasmodium falciparum transmission

    doi: 10.1186/s12936-017-1896-7

    Figure Lengend Snippet: Pfs25-specific serum antibody response after primary immunization with an Ad vector expressing Pfs25. Mice (n = 3 per group) were immunized with Ad5 expressing Pfs25 (10 10 particles) or recombinant yPfs25-alum (25 μg/mouse) on day 0, and serum samples were collected on day 21. a yPfs25-specific antibody titer. yPfs25-specific ELISA was performed and data depict pooled serum from 3 animals per group tested in triplicate in 3 independent ELISAs. The average ELISA unit and standard deviation are plotted. b yPfs25-specific avidity. Sera were added to a yPfs25-coated ELISA plate at a single dilution and then incubated with increasing concentrations of NaSCN. The remaining yPfs25-specific antibodies were detected with an HRP-conjugated anti-mouse IgG (Fc). The fraction of Ig bound was determined by dividing the optical density (OD) at each NaSCN concentration by the OD in the absence of NaSCN and the nonlinear best-fit line was plotted for each data set. c , d yPfs25-specific IgM and IgG subclass profile. yPfs25-specific ELISA was performed and the yPfs25-specific IgM and IgG subclasses were detected with HRP-conjugated specific antibodies. The average OD and standard deviation for each isotype/subclass ( c ) and the ratio of the O.D. for the IgG1 to the O.D.s of IgG2b and IgG3 ( d ) are shown

    Article Snippet: Briefly, the Pfs25 gene (bp 1–217) was codon-optimized for expression in humans (Genscript, Piscataway, NJ, USA) and inserted it into the pshuttle-CMV vector (Agilent Technologies plasmid #240007, Santa Clara, CA, USA).

    Techniques: Plasmid Preparation, Expressing, Mouse Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Standard Deviation, Incubation, Concentration Assay

    Plasmodium falciparum Pfs25 detection from RDT by direct real time-PCR. a DNA amplification kinetics per cycle on plate 1 for each sample. b logarithmic value of fluorescence and presence of background noise. c Amplification on plate 2, d DNA amplification at 8 cycles. Reactions performed on a conventional RT-PCR instrument (Bio-Rad CFX Connect) with a threshold setting of 50 relative fluorescence units (horizontal line)

    Journal: Malaria Journal

    Article Title: Molecular detection and quantification of Plasmodium falciparum gametocytes carriage in used RDTs in malaria elimination settings in northern Senegal

    doi: 10.1186/s12936-020-03204-w

    Figure Lengend Snippet: Plasmodium falciparum Pfs25 detection from RDT by direct real time-PCR. a DNA amplification kinetics per cycle on plate 1 for each sample. b logarithmic value of fluorescence and presence of background noise. c Amplification on plate 2, d DNA amplification at 8 cycles. Reactions performed on a conventional RT-PCR instrument (Bio-Rad CFX Connect) with a threshold setting of 50 relative fluorescence units (horizontal line)

    Article Snippet: Amplification of pfs25 gene by quantitative real-time PCR A qualitative and conventional qPCR protocol was standardized to detect pfs25 gene transcripts in RDT samples.

    Techniques: Real-time Polymerase Chain Reaction, Amplification, Fluorescence, Reverse Transcription Polymerase Chain Reaction

    Schematic diagram of the ‘launch’ vector. Following agroinfiltration of plants, the sequence between the left border (LB) and the right border (RB) of the plasmid vector is transferred from Agrobacteria into plant cells where expression of the engineered TMV genome is driven by the Cauliflower mosaic virus (CaMV) 35S promoter. TMV replicase then drives amplification of primary transcript, and Pfs25-CP accumulation is then driven by the TMV CP subgenomic promoter (light blue box). Movement protein (MP) facilitates cell-to-cell transfer of viral sequences and is driven by the MP subgenomic promoter (dark blue box).

    Journal: PLoS ONE

    Article Title: A Plant-Produced Pfs25 VLP Malaria Vaccine Candidate Induces Persistent Transmission Blocking Antibodies against Plasmodium falciparum in Immunized Mice

    doi: 10.1371/journal.pone.0079538

    Figure Lengend Snippet: Schematic diagram of the ‘launch’ vector. Following agroinfiltration of plants, the sequence between the left border (LB) and the right border (RB) of the plasmid vector is transferred from Agrobacteria into plant cells where expression of the engineered TMV genome is driven by the Cauliflower mosaic virus (CaMV) 35S promoter. TMV replicase then drives amplification of primary transcript, and Pfs25-CP accumulation is then driven by the TMV CP subgenomic promoter (light blue box). Movement protein (MP) facilitates cell-to-cell transfer of viral sequences and is driven by the MP subgenomic promoter (dark blue box).

    Article Snippet: The construct containing the Pfs25-CP sequence was optimized for plant expression by GeneArt (Regensburg, Germany) and cloned into the Tobacco mosaic virus (TMV)-based ‘launch’ expression vector pGR-D4 , .

    Techniques: Plasmid Preparation, Sequencing, Expressing, Amplification

    Anti-Pfs25 IgG responses in mice determined by ELISA. (A) Average anti-Pfs25 IgG titers from mice immunized with two doses of either 1.0 µg (circles) or 0.1 µg (squares) of Pfs25-CP VLPs, each with (open symbols) or without (filled symbols) Alhydrogel®, or 5.0 µg of CP only (black line). (B) Average anti-Pfs25 IgG titers from mice immunized with two doses of either 1.0 µg (circles), 0.1 µg (squares) or 0.01 µg (diamonds) of Pfs25-CP VLPs with Alhydrogel®. (C) Average anti-Pfs25 IgG titers elicited by a single administration of Pfs25-CP VLPs with Alhydrogel® at antigen doses ranging from 0.2–25 µg. Data are represented as average values per group of mice ± standard error of the mean.

    Journal: PLoS ONE

    Article Title: A Plant-Produced Pfs25 VLP Malaria Vaccine Candidate Induces Persistent Transmission Blocking Antibodies against Plasmodium falciparum in Immunized Mice

    doi: 10.1371/journal.pone.0079538

    Figure Lengend Snippet: Anti-Pfs25 IgG responses in mice determined by ELISA. (A) Average anti-Pfs25 IgG titers from mice immunized with two doses of either 1.0 µg (circles) or 0.1 µg (squares) of Pfs25-CP VLPs, each with (open symbols) or without (filled symbols) Alhydrogel®, or 5.0 µg of CP only (black line). (B) Average anti-Pfs25 IgG titers from mice immunized with two doses of either 1.0 µg (circles), 0.1 µg (squares) or 0.01 µg (diamonds) of Pfs25-CP VLPs with Alhydrogel®. (C) Average anti-Pfs25 IgG titers elicited by a single administration of Pfs25-CP VLPs with Alhydrogel® at antigen doses ranging from 0.2–25 µg. Data are represented as average values per group of mice ± standard error of the mean.

    Article Snippet: The construct containing the Pfs25-CP sequence was optimized for plant expression by GeneArt (Regensburg, Germany) and cloned into the Tobacco mosaic virus (TMV)-based ‘launch’ expression vector pGR-D4 , .

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Pfs25-CP VLP purity and identity. (A) Deduced amino acid sequence of Pfs25-CP with Pfs25 sequence underlined. Amino acids boxed in the sequence were identified by N-terminal sequencing of the SDS-PAGE bands indicated. A Coomassie stain of an SDS-PAGE gel highlights the Pfs25-CP fusion polypeptide (arrowhead ‘a’) and CP monomer polypeptides (arrowheads ‘b’ ‘c’). N-terminal sequencing of ‘a’ identified the first 5 amino acids of Pfs25, while sequencing of ‘b c’ identified residue 26 of AlMV CP. (B) Western blot analysis of Pfs25-CP VLPs with an anti-Pfs25 mAb (left panel) and an anti-AlMV CP polyclonal serum (right panel).

    Journal: PLoS ONE

    Article Title: A Plant-Produced Pfs25 VLP Malaria Vaccine Candidate Induces Persistent Transmission Blocking Antibodies against Plasmodium falciparum in Immunized Mice

    doi: 10.1371/journal.pone.0079538

    Figure Lengend Snippet: Pfs25-CP VLP purity and identity. (A) Deduced amino acid sequence of Pfs25-CP with Pfs25 sequence underlined. Amino acids boxed in the sequence were identified by N-terminal sequencing of the SDS-PAGE bands indicated. A Coomassie stain of an SDS-PAGE gel highlights the Pfs25-CP fusion polypeptide (arrowhead ‘a’) and CP monomer polypeptides (arrowheads ‘b’ ‘c’). N-terminal sequencing of ‘a’ identified the first 5 amino acids of Pfs25, while sequencing of ‘b c’ identified residue 26 of AlMV CP. (B) Western blot analysis of Pfs25-CP VLPs with an anti-Pfs25 mAb (left panel) and an anti-AlMV CP polyclonal serum (right panel).

    Article Snippet: The construct containing the Pfs25-CP sequence was optimized for plant expression by GeneArt (Regensburg, Germany) and cloned into the Tobacco mosaic virus (TMV)-based ‘launch’ expression vector pGR-D4 , .

    Techniques: Sequencing, SDS Page, Staining, Western Blot

    Pfs25-CP VLP particle analysis. (A) Negative stain transmission electron micrograph of Pfs25-CP VLPs shows highly uniform particles of 19.3±2.4 nm in diameter. (B) Transmission electron micrograph of Pfs25-CP VLPs labeled with anti-Pfs25 and gold-labeled anti-mouse antibodies confirms the presence of Pfs25 on the particles. (C) DLS histogram showing a narrow size distribution for Pfs25-CP VLPs. The average hydrodynamic radius is ∼14 nm with a polydispersity of

    Journal: PLoS ONE

    Article Title: A Plant-Produced Pfs25 VLP Malaria Vaccine Candidate Induces Persistent Transmission Blocking Antibodies against Plasmodium falciparum in Immunized Mice

    doi: 10.1371/journal.pone.0079538

    Figure Lengend Snippet: Pfs25-CP VLP particle analysis. (A) Negative stain transmission electron micrograph of Pfs25-CP VLPs shows highly uniform particles of 19.3±2.4 nm in diameter. (B) Transmission electron micrograph of Pfs25-CP VLPs labeled with anti-Pfs25 and gold-labeled anti-mouse antibodies confirms the presence of Pfs25 on the particles. (C) DLS histogram showing a narrow size distribution for Pfs25-CP VLPs. The average hydrodynamic radius is ∼14 nm with a polydispersity of

    Article Snippet: The construct containing the Pfs25-CP sequence was optimized for plant expression by GeneArt (Regensburg, Germany) and cloned into the Tobacco mosaic virus (TMV)-based ‘launch’ expression vector pGR-D4 , .

    Techniques: Staining, Transmission Assay, Labeling

    The effect of TSA on P. falciparum sexual stage development. (A) The effect of TSA on gametocyte development. TSA at IC 50 or IC 90 concentrations was added to stage II gametocyte cultures for 2 days. The numbers of stage IV and V gametocytes were determined in 1,000 erythrocytes at day 10 using Giemsa-stained blood smears. Epoxomicin (60 nM) was used as a positive control (not shown), while 0.5% vol. ethanol and chloroquine (16 nM) were used as negative controls (ethanol set to 100%). (B) The effect of TSA on macrogamete development. A mature gametocyte culture was incubated with TSA at IC 50 or IC 90 concentrations or 0.5% vol. ethanol for 1 h at 37°C. The culture was then activated with 100 μM XA and further cultured for 30 min at RT for macrogamete development. Macrogametes were detected by immunolabelling with anti-Pfs25 and counted in triplicate in 1,000 erythrocytes. (C) Effect of TSA on zygote development. The zygote development assay was performed in the same way as the macrogamete development except for the fact that after activation the cultures were incubated for 12 h at RT. Results shown (for A–C ) are combined from three independent experiments each performed in triplicate (mean ± SD). * P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Transcriptional Profiling Defines Histone Acetylation as a Regulator of Gene Expression during Human-to-Mosquito Transmission of the Malaria Parasite Plasmodium falciparum

    doi: 10.3389/fcimb.2017.00320

    Figure Lengend Snippet: The effect of TSA on P. falciparum sexual stage development. (A) The effect of TSA on gametocyte development. TSA at IC 50 or IC 90 concentrations was added to stage II gametocyte cultures for 2 days. The numbers of stage IV and V gametocytes were determined in 1,000 erythrocytes at day 10 using Giemsa-stained blood smears. Epoxomicin (60 nM) was used as a positive control (not shown), while 0.5% vol. ethanol and chloroquine (16 nM) were used as negative controls (ethanol set to 100%). (B) The effect of TSA on macrogamete development. A mature gametocyte culture was incubated with TSA at IC 50 or IC 90 concentrations or 0.5% vol. ethanol for 1 h at 37°C. The culture was then activated with 100 μM XA and further cultured for 30 min at RT for macrogamete development. Macrogametes were detected by immunolabelling with anti-Pfs25 and counted in triplicate in 1,000 erythrocytes. (C) Effect of TSA on zygote development. The zygote development assay was performed in the same way as the macrogamete development except for the fact that after activation the cultures were incubated for 12 h at RT. Results shown (for A–C ) are combined from three independent experiments each performed in triplicate (mean ± SD). * P

    Article Snippet: Antibodies Primary antibodies used in this study included: rabbit anti-(tetra)-acetyl histone H4 K5, 8, 12, 16ac (H4Kac4) (Millipore; note: according the manufacturer's material data sheet this antibody may cross-react with other acetylated histones like H2B); rabbit anti-H3K9ac (Diagenode); rabbit anti-PfHP1 (Brancucci et al., ); rabbit IgG antibody (Millipore); mouse/rabbit anti-Pfs230 (Ngwa et al., ; Simon et al., ), rabbit anti-Pfs25 (BEI Resources); mouse anti-Pf39 (Scholz et al., ); rabbit anti-HA (Sigma Aldrich); mouse anti-proteasome SU α5 (Aminake et al., ), and anti-PfActinI (Ngwa et al., ).

    Techniques: Staining, Positive Control, Incubation, Cell Culture, Activation Assay

    SDS-PAGE and Western blot (4B7) of Pichia and baculovirus Pfs25. a SDS-PAGE and b Western blot using 4B7 monoclonal antibody of purified Pfs25 from Pichia under reducing conditions (non-reduced not show). c SDS-PAGE and d Western blot using 4B7 monoclonal antibody of non-reduced and reduced purified Pfs25 from baculovirus

    Journal: Malaria Journal

    Article Title: Assessment of Pfs25 expressed from multiple soluble expression platforms for use as transmission-blocking vaccine candidates

    doi: 10.1186/s12936-016-1464-6

    Figure Lengend Snippet: SDS-PAGE and Western blot (4B7) of Pichia and baculovirus Pfs25. a SDS-PAGE and b Western blot using 4B7 monoclonal antibody of purified Pfs25 from Pichia under reducing conditions (non-reduced not show). c SDS-PAGE and d Western blot using 4B7 monoclonal antibody of non-reduced and reduced purified Pfs25 from baculovirus

    Article Snippet: Three high-yield clones were identified by analyses with reducing SDS-PAGE (Pfs25 expression band at 20 kDa), and evaluated with anti-His (Qiagen) or anti-Pfs25 mAb 4B7 [ , , ] (BEI Resources) Western blots as described.

    Techniques: SDS Page, Western Blot, Purification

    Characterization of Pfs25-EPA conjugate desorbed from Alhydrogel formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.

    Journal: Vaccine

    Article Title: Accelerated and long term stability study of Pfs25-EPA conjugates adjuvanted with Alhydrogel®

    doi: 10.1016/j.vaccine.2017.04.067

    Figure Lengend Snippet: Characterization of Pfs25-EPA conjugate desorbed from Alhydrogel formulation following 4 years storage at 2-8°C A) SDS-PAGE (silver staining); B) Western blot: (a) probed with monoclonal antibody 4B7; (b) probed with penta-His; (c) probed with anti-Exotoxin A. Lane 1, Pfs25-EPA reference standard (without desorption); Lane 2, Pfs25-EPA/Alhydrogel desorption. All samples were loaded at 550 ng protein (calculation based on 100% recovery of desorption) per lane. Molecular weight markers are indicated in kDa.

    Article Snippet: The general safety analysis for clinical grade Pfs25-EPA/Alhydrogel vaccine was performed by BioReliance (Rockville, Maryland) per 21 CFR, Section 610.11 and USP Chapter < 88 > .

    Techniques: SDS Page, Silver Staining, Western Blot, Molecular Weight