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LakePharma 3c1 igg1 igg3
Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs <t>2H8-IgG1/IgG2a</t> and <t>3C1-IgG1/IgG3</t> were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea
3c1 Igg1 Igg3, supplied by LakePharma, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children"

Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

Journal: BMC Medicine

doi: 10.1186/s12916-018-1054-2

Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea
Figure Legend Snippet: Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea

Techniques Used: Peptide ELISA, Enzyme-linked Immunosorbent Assay

Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2
Figure Legend Snippet: Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2

Techniques Used: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea
Figure Legend Snippet: Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea

Techniques Used: Enzyme-linked Immunosorbent Assay

Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization
Figure Legend Snippet: Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization

Techniques Used: Purification, Enzyme-linked Immunosorbent Assay

Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization
Figure Legend Snippet: Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization

Techniques Used: Inhibition, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation, In Vitro

2) Product Images from "Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children"

Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

Journal: BMC Medicine

doi: 10.1186/s12916-018-1054-2

Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea
Figure Legend Snippet: Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea

Techniques Used: Peptide ELISA, Enzyme-linked Immunosorbent Assay

Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2
Figure Legend Snippet: Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2

Techniques Used: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea
Figure Legend Snippet: Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea

Techniques Used: Enzyme-linked Immunosorbent Assay

Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization
Figure Legend Snippet: Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization

Techniques Used: Purification, Enzyme-linked Immunosorbent Assay

Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization
Figure Legend Snippet: Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization

Techniques Used: Inhibition, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation, In Vitro

Naturally acquired human anti-CSP IgG correlates with the ability to promote C1q-fixation to CSP, despite individual differences in reactivity. Antibodies from malaria-exposed adults living in PNG ( a ; N = 116) and Kenya ( b ; N = 104) were tested for IgG and C1q-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean and range of duplicates were graphed (mean only for scatter plots). The left panels show correlations between IgG and C1q-fixation to CSP (Spearman’s correlation coefficient, rho). The right panels show selected examples of IgG and C1q-fixation to CSP for individual serum samples from PNG donors (V7, V49, V51, V52, and V53) and Kenyan donors (AR18, AR22, AR28, AR36, and AR47). AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, PNG Papua New Guinea
Figure Legend Snippet: Naturally acquired human anti-CSP IgG correlates with the ability to promote C1q-fixation to CSP, despite individual differences in reactivity. Antibodies from malaria-exposed adults living in PNG ( a ; N = 116) and Kenya ( b ; N = 104) were tested for IgG and C1q-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean and range of duplicates were graphed (mean only for scatter plots). The left panels show correlations between IgG and C1q-fixation to CSP (Spearman’s correlation coefficient, rho). The right panels show selected examples of IgG and C1q-fixation to CSP for individual serum samples from PNG donors (V7, V49, V51, V52, and V53) and Kenyan donors (AR18, AR22, AR28, AR36, and AR47). AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, PNG Papua New Guinea

Techniques Used: Enzyme-linked Immunosorbent Assay

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    LakePharma 3c1 igg1 igg3
    Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs <t>2H8-IgG1/IgG2a</t> and <t>3C1-IgG1/IgG3</t> were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea
    3c1 Igg1 Igg3, supplied by LakePharma, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Peptide ELISA, Enzyme-linked Immunosorbent Assay

    Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Enzyme-linked Immunosorbent Assay

    Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Purification, Enzyme-linked Immunosorbent Assay

    Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation, In Vitro

    Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Antibodies that target the NANP epitope of CSP can promote C1q-fixation. Antibodies from malaria-exposed adults living in PNG ( n = 30) ( a ) and Kenya ( n = 30) ( b ) were tested for IgG, IgM, and C1q-fixation to (NANP) 15 peptide by ELISA, and correlated with C1q fixation to CSP (Spearman’s correlation coefficient, rho). Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines) and mean of duplicates were graphed. c Mouse anti-CSP MAbs 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 were tested for CSP-IgG, NANP-IgG, and C1q-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of duplicates were graphed. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, IgM immunoglobulin M, MAb monoclonal antibody, OD optical density, PNG Papua New Guinea

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Peptide ELISA, Enzyme-linked Immunosorbent Assay

    Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Acquisition of C1q-fixing antibodies and total IgG to CSP and MSP2 antigens in malaria-exposed Kenyan children and adults. Kenyan children and adults ( N = 75) were categorized into groups based on the median ages 0.5, 1, 2, 5, and 35 years ( n = 11, n = 14, n = 18, n = 16, and n = 16, respectively). Samples were tested for C1q-fixation and total IgG to CSP ( a ) and MSP2 ( b ) by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naive negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean of duplicates were graphed along with the group median, interquartile range, and percentage of positive samples. Reactivity between two groups and more than two groups were compared using the Mann–Whitney U test and Kruskal–Wallis test, respectively. AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, MSP2 Merozoite surface protein 2

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Naturally acquired human anti-CSP antibodies are predominately IgG1, IgG3, and IgM, and can promote complement fixation to CSP. Antibodies from malaria-exposed adults ( n = 30 in each group) living in PNG ( a , c-e ) and Kenyan ( b ) were tested for IgG/IgM and complement-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and mean and range of duplicates were graphed (mean only for scatter plots). a,b IgG subclasses and IgM reactivity to CSP. The median, interquartile range, and percentage of positive samples are shown. c Correlations between C1q-fixation and C4/C3/C3/C5b-C9-fixation to CSP (Spearman’s correlation coefficient, rho). d Examples of C1q and C3-fixation to CSP by individual serum samples (V15, V32, V33, V45, and V46 from PNG donors, n = 5). e C3-fixation to CSP by individual samples (V6, V7, V32, V37, and V43 from PNG donors, n = 5) in the presence of normal human serum (NHS) and serum depleted of C1q (C1q dep.) (Wilcoxon matched-pairs signed rank test). Non-standardized data are shown, as only one Melbourne control (Melb.) was tested. AU arbitrary units, CSP circumsporozoite protein, dep. depleted, ELISA enzyme-linked immunosorbent assay, Melb. Melbourne, NHS normal human serum, PNG Papua New Guinea

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Enzyme-linked Immunosorbent Assay

    Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Vaccine-induced rabbit anti-CSP IgG can fix human complement proteins to CSP. Purified IgG from rabbit serum before (pre-imm.) and after (a-CSP) CSP immunization was tested for IgG and complement-fixation to CSP by ELISA. Results were corrected for background reactivity using no-IgG controls, and the mean and range of the duplicates were graphed. a IgG reactivity to CSP and (NANP) 15 peptide (pre-immune IgG shown with the open symbol was tested at 10 μg/ml). Results from two independent experiments are shown. b C1q and C3-fixation to CSP tested in the presence (+) and absence (−) of complement to confirm specificity for complement fixation. a-CSP after CSP immunization, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, pre-imm. before CSP immunization

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Purification, Enzyme-linked Immunosorbent Assay

    Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Antibodies promote complement-fixation on P. falciparum sporozoites, which enhances antibody-mediated traversal inhibition and can lead to sporozoite death. a Antibody samples from malaria-exposed (PNG and Kenyan) and malaria-naïve (Melbourne) individuals were tested for the ability to fix human C1q and C3 to 3D7 P. falciparum sporozoites by ELISA and Western blot. ELISA samples (top panel) were tested in duplicate, and the mean and range were graphed (C1q-fixation data is from two independent experiments). Western blot sporozoites (bottom panel) were incubated with human antibody samples and normal human serum (NHS, active complement), and then washed and processed for Western blotting under reduced conditions. Any complement proteins that had deposited on the sporozoite surface were detected using C1q- and C3-specific antibodies, and the sporozoite surface antigen, CSP, was used as a loading control. b,c In vitro traversal inhibition of freshly dissected sporozoites incubated with HC-04 cells, in the presence of NHS and heat-inactivated human serum (HIS). Each condition was tested in duplicate, and the mean and range were graphed. b Traversal inhibition of NF54 sporozoites treated with rabbit anti-CSP IgG (compared to rabbit pre-immune IgG). c Traversal inhibition of NF54 and NF166.C8 sporozoites treated with malaria-exposed Kenyan pool (compared to malaria-naïve Melbourne pool). Results from two independent experiments are shown. d Percentage of dead 3D7 sporozoites (PI+ cells) treated with rabbit anti-CSP IgG and pre-immune IgG, in the presence of NHS or C5-depleted serum (C5dep.). The mean and range of two independent experiments are graphed. a-CSP after CSP immunization, C5dep. C5-depleted serum, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, HIS heat-inactivated human serum, IgG immunoglobulin G, Melb. Melbourne, NHS normal human serum, OD optical density, PI propidium iodide, PNG Papua New Guinea, pre-imm. before CSP immunization

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation, In Vitro

    Naturally acquired human anti-CSP IgG correlates with the ability to promote C1q-fixation to CSP, despite individual differences in reactivity. Antibodies from malaria-exposed adults living in PNG ( a ; N = 116) and Kenya ( b ; N = 104) were tested for IgG and C1q-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean and range of duplicates were graphed (mean only for scatter plots). The left panels show correlations between IgG and C1q-fixation to CSP (Spearman’s correlation coefficient, rho). The right panels show selected examples of IgG and C1q-fixation to CSP for individual serum samples from PNG donors (V7, V49, V51, V52, and V53) and Kenyan donors (AR18, AR22, AR28, AR36, and AR47). AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, PNG Papua New Guinea

    Journal: BMC Medicine

    Article Title: Human antibodies activate complement against Plasmodium falciparum sporozoites, and are associated with protection against malaria in children

    doi: 10.1186/s12916-018-1054-2

    Figure Lengend Snippet: Naturally acquired human anti-CSP IgG correlates with the ability to promote C1q-fixation to CSP, despite individual differences in reactivity. Antibodies from malaria-exposed adults living in PNG ( a ; N = 116) and Kenya ( b ; N = 104) were tested for IgG and C1q-fixation to CSP by ELISA. Results were standardized to arbitrary units (AU) based on malaria-naïve negative controls from Melbourne (seropositivity defined as AU > 1, shown as dotted lines), and the mean and range of duplicates were graphed (mean only for scatter plots). The left panels show correlations between IgG and C1q-fixation to CSP (Spearman’s correlation coefficient, rho). The right panels show selected examples of IgG and C1q-fixation to CSP for individual serum samples from PNG donors (V7, V49, V51, V52, and V53) and Kenyan donors (AR18, AR22, AR28, AR36, and AR47). AU arbitrary units, CSP circumsporozoite protein, ELISA enzyme-linked immunosorbent assay, IgG immunoglobulin G, PNG Papua New Guinea

    Article Snippet: We were provided with subclass switched mouse anti-NANP monoclonal antibodies (MAbs) 2H8-IgG1/IgG2a and 3C1-IgG1/IgG3 (LakePharma, Belmont, USA) by PATH’s MVI [ ].

    Techniques: Enzyme-linked Immunosorbent Assay