glycoengineered anti cd20 mabs  (Sino Biological)


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    Name:
    Anti CD20 Antibody Mouse Monoclonal
    Description:
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant Human CD20 MS4A1 rh CD20 MS4A1 Catalog 11007 H07H2 NP 068769 2 Glu213 Pro297 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    Catalog Number:
    11007-mm01
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    Human
    Applications:
    ELISA
    Immunogen:
    Recombinant Human CD20 / MS4A1 Protein (Catalog#11007-H07H2)
    Product Aliases:
    Anti-B1 Antibody, Anti-Bp35 Antibody, Anti-CD20 Antibody, Anti-CVID5 Antibody, Anti-LEU-16 Antibody, Anti-MS4A1 Antibody, Anti-MS4A2 Antibody, Anti-S7 Antibody
    Antibody Type:
    MAb
    Host:
    Mouse
    Isotype:
    Mouse IgG2b
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    Structured Review

    Sino Biological glycoengineered anti cd20 mabs
    FcγR activation properties of <t>glycoengineered</t> <t>anti-CD20</t> mAbs. FcγRIIa (a) and FcγRIIIa (b) activation properties of glycoengineered mAbs. Jurkat/FcγRIIa/NFAT-Luc or Jurkat/FcγRIIIa/NFAT-Luc reporter cells were incubated with serially diluted anti-CD20 mAbs in the presence of Raji cells. FcγR activation was evaluated by assessing the luminescence intensity.
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant Human CD20 MS4A1 rh CD20 MS4A1 Catalog 11007 H07H2 NP 068769 2 Glu213 Pro297 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    https://www.bioz.com/result/glycoengineered anti cd20 mabs/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    glycoengineered anti cd20 mabs - by Bioz Stars, 2021-03
    92/100 stars

    Images

    1) Product Images from "Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies"

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies

    Journal: mAbs

    doi: 10.1080/19420862.2019.1608143

    FcγR activation properties of glycoengineered anti-CD20 mAbs. FcγRIIa (a) and FcγRIIIa (b) activation properties of glycoengineered mAbs. Jurkat/FcγRIIa/NFAT-Luc or Jurkat/FcγRIIIa/NFAT-Luc reporter cells were incubated with serially diluted anti-CD20 mAbs in the presence of Raji cells. FcγR activation was evaluated by assessing the luminescence intensity.
    Figure Legend Snippet: FcγR activation properties of glycoengineered anti-CD20 mAbs. FcγRIIa (a) and FcγRIIIa (b) activation properties of glycoengineered mAbs. Jurkat/FcγRIIa/NFAT-Luc or Jurkat/FcγRIIIa/NFAT-Luc reporter cells were incubated with serially diluted anti-CD20 mAbs in the presence of Raji cells. FcγR activation was evaluated by assessing the luminescence intensity.

    Techniques Used: Activation Assay, Incubation

    CDC and C1q-binding activities of glycoengineered anti-CD20 mAbs. (a, b) CDC activities of glycoengineered anti-CD20 mAbs. Raji cells were incubated with 16% human serum and were serially diluted with glycoengineered anti-CD20 mAbs. (a) Percentages of cell lysis plotted against mAb concentrations. (b) CDC activity of 1 µg/ml of G1aF mAb and G1bF mAb mixtures in different ratios. (c) C1q binding of glycoengineered anti-CD20 mAbs. Raji cells were opsonized with anti-CD20 mAbs and incubated with human serum. The cells were stained with FITC-conjugated anti-C1q antibody and the C1q-binding level was analyzed by flow cytometer. Data are presented as mean ± SD ( n = 3).
    Figure Legend Snippet: CDC and C1q-binding activities of glycoengineered anti-CD20 mAbs. (a, b) CDC activities of glycoengineered anti-CD20 mAbs. Raji cells were incubated with 16% human serum and were serially diluted with glycoengineered anti-CD20 mAbs. (a) Percentages of cell lysis plotted against mAb concentrations. (b) CDC activity of 1 µg/ml of G1aF mAb and G1bF mAb mixtures in different ratios. (c) C1q binding of glycoengineered anti-CD20 mAbs. Raji cells were opsonized with anti-CD20 mAbs and incubated with human serum. The cells were stained with FITC-conjugated anti-C1q antibody and the C1q-binding level was analyzed by flow cytometer. Data are presented as mean ± SD ( n = 3).

    Techniques Used: Binding Assay, Incubation, Lysis, Activity Assay, Staining, Flow Cytometry

    Comparison of structural stabilities of the CH2 domain of glycoengineered anti-CD20 mAbs using HDX/MS . Deuterium uptake plots of peptides Phe245–Leu255 (FLFPPKPKDTL) (a), Leu246–Leu255 (LFPPKPKDTL) (b), and Leu246–Met256 (LFPPKPKDTLM) (c) in the H-chain. (d) Physical representations of the crystal structures (PDB 1HZH) of peptides at Phe245–Met256 (magenta ribbons).
    Figure Legend Snippet: Comparison of structural stabilities of the CH2 domain of glycoengineered anti-CD20 mAbs using HDX/MS . Deuterium uptake plots of peptides Phe245–Leu255 (FLFPPKPKDTL) (a), Leu246–Leu255 (LFPPKPKDTL) (b), and Leu246–Met256 (LFPPKPKDTLM) (c) in the H-chain. (d) Physical representations of the crystal structures (PDB 1HZH) of peptides at Phe245–Met256 (magenta ribbons).

    Techniques Used:

    Glycan profiles of mAbs with homogeneous glycans. Glycan profiles of commercially available anti-CD20 mAb (a) and glycoengineered G0F mAb (b), G1aF mAb (c), G1bF mAb (d), and G2F mAb (e). The glycan profiles were obtained using HPLC analysis of 2-AB-labeled glycans.
    Figure Legend Snippet: Glycan profiles of mAbs with homogeneous glycans. Glycan profiles of commercially available anti-CD20 mAb (a) and glycoengineered G0F mAb (b), G1aF mAb (c), G1bF mAb (d), and G2F mAb (e). The glycan profiles were obtained using HPLC analysis of 2-AB-labeled glycans.

    Techniques Used: High Performance Liquid Chromatography, Labeling

    Binding of glycoengineered anti-CD20 mAbs to human FcγRs. SPR analysis was used to measure the binding of anti-CD20 mAbs to human FcγRI, FcγRIIa, and FcγRIIIa. Binding sensorgrams corrected by both the surface blank and buffer injection control are represented.
    Figure Legend Snippet: Binding of glycoengineered anti-CD20 mAbs to human FcγRs. SPR analysis was used to measure the binding of anti-CD20 mAbs to human FcγRI, FcγRIIa, and FcγRIIIa. Binding sensorgrams corrected by both the surface blank and buffer injection control are represented.

    Techniques Used: Binding Assay, SPR Assay, Injection

    2) Product Images from "Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)"

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    Journal: mAbs

    doi: 10.1080/19420862.2015.1078054

    Expression of anti-CD20 mAb in transgenic silkworms. ( A ) The protein lysates extracted from MSGs of H line or L line transgenic silkworms were separated by SDS-PAGE followed by staining with CBB or by western blotting with an anti-Human IgG(H + L) antibody. The protein lysates extracted form MSGs of transgenic silkworms that harbored only the Ser1-GAL4 construct were used as negative controls. The numbers above the gels or western blots indicate the line number of each transgenic strain. ( B ) The protein lysates extracted form MSGs or cocoons of the H + L line transgenic silkworms were separated by SDS-PAGE under reducing or non-reducing conditions. Gels were stained with CBB or subjected to protein gel blotting using an anti-Human IgG(H + L) antibody. The lysates obtained from the transgenic silkworms harboring only the Ser1-GAL4 construct were used as a negative control.
    Figure Legend Snippet: Expression of anti-CD20 mAb in transgenic silkworms. ( A ) The protein lysates extracted from MSGs of H line or L line transgenic silkworms were separated by SDS-PAGE followed by staining with CBB or by western blotting with an anti-Human IgG(H + L) antibody. The protein lysates extracted form MSGs of transgenic silkworms that harbored only the Ser1-GAL4 construct were used as negative controls. The numbers above the gels or western blots indicate the line number of each transgenic strain. ( B ) The protein lysates extracted form MSGs or cocoons of the H + L line transgenic silkworms were separated by SDS-PAGE under reducing or non-reducing conditions. Gels were stained with CBB or subjected to protein gel blotting using an anti-Human IgG(H + L) antibody. The lysates obtained from the transgenic silkworms harboring only the Ser1-GAL4 construct were used as a negative control.

    Techniques Used: Expressing, Transgenic Assay, SDS Page, Staining, Western Blot, Construct, Negative Control

    FcγRIIIa activation and antibody-dependent cell-mediated cytotoxicity (ADCC). ( A ) CD20 binding-dependent activation of FcγRIIIa was measured by using Daudi (target) and Jurkat/FcγRIIIa/NFAT-Luc (effector) cells. Fold increases in luciferase activity were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) The levels of ADCC against Daudi cells were measured by using human PBMCs as effector cells. The percentages of cell lysis were plotted against the mAbs concentration (n = 3, bars indicate SEM).
    Figure Legend Snippet: FcγRIIIa activation and antibody-dependent cell-mediated cytotoxicity (ADCC). ( A ) CD20 binding-dependent activation of FcγRIIIa was measured by using Daudi (target) and Jurkat/FcγRIIIa/NFAT-Luc (effector) cells. Fold increases in luciferase activity were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) The levels of ADCC against Daudi cells were measured by using human PBMCs as effector cells. The percentages of cell lysis were plotted against the mAbs concentration (n = 3, bars indicate SEM).

    Techniques Used: Activation Assay, Binding Assay, Luciferase, Activity Assay, Concentration Assay, Lysis

    CD20-binding properties. ( A ) Daudi cells were incubated with anti-CD20 mAbs and their binding properties were assessed by flow cytometric analysis using Alexa488-conjugated F(ab')2 anti-human IgG Fc. The mean fluorescence intensities were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) Daudi cells were incubated with DyLight 488-labeled MabThera and serially diluted unlabeled mAbs. Percentages of DyLight 488-labeled MabThera binding were calculated by the mean fluorescence intensities and plotted against the unlabeled mAbs concentration (n = 3, bars indicate SEM).
    Figure Legend Snippet: CD20-binding properties. ( A ) Daudi cells were incubated with anti-CD20 mAbs and their binding properties were assessed by flow cytometric analysis using Alexa488-conjugated F(ab')2 anti-human IgG Fc. The mean fluorescence intensities were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) Daudi cells were incubated with DyLight 488-labeled MabThera and serially diluted unlabeled mAbs. Percentages of DyLight 488-labeled MabThera binding were calculated by the mean fluorescence intensities and plotted against the unlabeled mAbs concentration (n = 3, bars indicate SEM).

    Techniques Used: Binding Assay, Incubation, Fluorescence, Concentration Assay, Labeling

    Complement-dependent cytotoxicity (CDC). Daudi cells were cultured in the presence of human serum (16%) and anti-CD20 mAbs (1, 3 or 10 µg/ml). The percentages of 7-AAD positive-dead cells were calculated by flow cytometric analysis and represented as the mean + SEM (n = 3). **, p
    Figure Legend Snippet: Complement-dependent cytotoxicity (CDC). Daudi cells were cultured in the presence of human serum (16%) and anti-CD20 mAbs (1, 3 or 10 µg/ml). The percentages of 7-AAD positive-dead cells were calculated by flow cytometric analysis and represented as the mean + SEM (n = 3). **, p

    Techniques Used: Cell Culture

    Structures of the plasmids used to generate transgenic silkworms. Each plasmid has right and left arms of piggyBac and the 3 × P3-fluorescent gene cassette for a screening marker (EYFP, AmCyan, or DsRed2). Plasmids pBac[UAS_anti-CD20 mAb HC/3 × P3-EYFP] and pBac[UAS_anti-CD20 mAb LC/3 × P3-AmCyan] encode the anti-CD20 mAb H chain gene and the anti-CD20 mAb L chain gene, respectively, under the control of a UAS promoter, and contain an BmNPV- derived hr5 enhancer and an A3-Blasticidin cassette. The anti-CD20 H and L genes were fused to the signal peptide sequence of the sericin 1 gene encoded by exon 1 and 18 bp of exon 2. The plasmid pBac[Ser1-GAL4/3 × P3-DsRed] encodes the GAL4 gene under the control of the sericin1 promoter.
    Figure Legend Snippet: Structures of the plasmids used to generate transgenic silkworms. Each plasmid has right and left arms of piggyBac and the 3 × P3-fluorescent gene cassette for a screening marker (EYFP, AmCyan, or DsRed2). Plasmids pBac[UAS_anti-CD20 mAb HC/3 × P3-EYFP] and pBac[UAS_anti-CD20 mAb LC/3 × P3-AmCyan] encode the anti-CD20 mAb H chain gene and the anti-CD20 mAb L chain gene, respectively, under the control of a UAS promoter, and contain an BmNPV- derived hr5 enhancer and an A3-Blasticidin cassette. The anti-CD20 H and L genes were fused to the signal peptide sequence of the sericin 1 gene encoded by exon 1 and 18 bp of exon 2. The plasmid pBac[Ser1-GAL4/3 × P3-DsRed] encodes the GAL4 gene under the control of the sericin1 promoter.

    Techniques Used: Transgenic Assay, Plasmid Preparation, Marker, Derivative Assay, Sequencing

    Purification of anti-CD20 mAbs from the transgenic silkworms. ( A ) Anti-CD20 mAbs were purified from the lysates of MSGs or cocoons derived from transgenic silkworms by affinity chromatography using a Protein G column. The lysates (original), flow-through fractions from the Protein G column, and eluted fractions were analyzed by non-reducing SDS-PAGE. ( B ) MabThera and anti-CD20 mAbs derived from transgenic silkworms showed the same migration pattern on non-reducing SDS-PAGE.
    Figure Legend Snippet: Purification of anti-CD20 mAbs from the transgenic silkworms. ( A ) Anti-CD20 mAbs were purified from the lysates of MSGs or cocoons derived from transgenic silkworms by affinity chromatography using a Protein G column. The lysates (original), flow-through fractions from the Protein G column, and eluted fractions were analyzed by non-reducing SDS-PAGE. ( B ) MabThera and anti-CD20 mAbs derived from transgenic silkworms showed the same migration pattern on non-reducing SDS-PAGE.

    Techniques Used: Purification, Transgenic Assay, Derivative Assay, Affinity Chromatography, SDS Page, Migration

    Fc glycosylation analysis of anti-CD20 mAbs. ( A ) Comparison of mass spectra acquired at the elution point of the glycopeptides of EEQYNSTYR, corresponding to 293–301 amino acids (EU numbering) in the heavy chain of anti-CD20 mAbs. ( B ) Percentage distributions of glycoforms were calculated by the relative peak area average values. Symbols: blue square, N-acetylglucosamine; green circle, mannose; yellow circle, galactose; red triangle, fucose.
    Figure Legend Snippet: Fc glycosylation analysis of anti-CD20 mAbs. ( A ) Comparison of mass spectra acquired at the elution point of the glycopeptides of EEQYNSTYR, corresponding to 293–301 amino acids (EU numbering) in the heavy chain of anti-CD20 mAbs. ( B ) Percentage distributions of glycoforms were calculated by the relative peak area average values. Symbols: blue square, N-acetylglucosamine; green circle, mannose; yellow circle, galactose; red triangle, fucose.

    Techniques Used:

    Binding affinity to human Fc receptors. Binding affinities of anti-CD20 mAbs to FcγRI, FcγRIIIa and FcRn were measured by an SPR analysis. ( A ) Binding sensorgrams corrected for both the surface blank and the buffer injection control are represented. Dashed lines indicate the fitting curves generated by 1:1 binding model for FcγRs and bivalent model for FcRn, respectively. ( B ) Dissociation constant (K D ) values are represented as mean + SD (n = 3). *, p
    Figure Legend Snippet: Binding affinity to human Fc receptors. Binding affinities of anti-CD20 mAbs to FcγRI, FcγRIIIa and FcRn were measured by an SPR analysis. ( A ) Binding sensorgrams corrected for both the surface blank and the buffer injection control are represented. Dashed lines indicate the fitting curves generated by 1:1 binding model for FcγRs and bivalent model for FcRn, respectively. ( B ) Dissociation constant (K D ) values are represented as mean + SD (n = 3). *, p

    Techniques Used: Binding Assay, SPR Assay, Injection, Generated

    Related Articles

    Injection:

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)
    Article Snippet: For measurement of the binding affinity with FcγRs, recombinant ectodomains of humnan FcγRs with a C-terminal polyhistidine tags (Sino Biological) were captured on an anti-polyhistidine antibody-immobilized sensorchip. .. Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored. .. The dissociation constant K D was calculated using the 1 : 1 binding model.

    Binding Assay:

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies
    Article Snippet: Surface plasmon resonance analysis A Biacore T200 SPR biosensor (GE Healthcare) and CM5 sensor chip were used to evaluate the binding properties of glycoengineered anti-CD20 mAbs. .. The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously. .. In all experiments, each sensorgram from the sample flow cell was corrected by both surface blank and buffer injection control (double reference).

    Recombinant:

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies
    Article Snippet: Surface plasmon resonance analysis A Biacore T200 SPR biosensor (GE Healthcare) and CM5 sensor chip were used to evaluate the binding properties of glycoengineered anti-CD20 mAbs. .. The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously. .. In all experiments, each sensorgram from the sample flow cell was corrected by both surface blank and buffer injection control (double reference).

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    Sino Biological glycoengineered anti cd20 mabs
    FcγR activation properties of <t>glycoengineered</t> <t>anti-CD20</t> mAbs. FcγRIIa (a) and FcγRIIIa (b) activation properties of glycoengineered mAbs. Jurkat/FcγRIIa/NFAT-Luc or Jurkat/FcγRIIIa/NFAT-Luc reporter cells were incubated with serially diluted anti-CD20 mAbs in the presence of Raji cells. FcγR activation was evaluated by assessing the luminescence intensity.
    Glycoengineered Anti Cd20 Mabs, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glycoengineered anti cd20 mabs/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    glycoengineered anti cd20 mabs - by Bioz Stars, 2021-03
    92/100 stars
      Buy from Supplier

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    FcγR activation properties of glycoengineered anti-CD20 mAbs. FcγRIIa (a) and FcγRIIIa (b) activation properties of glycoengineered mAbs. Jurkat/FcγRIIa/NFAT-Luc or Jurkat/FcγRIIIa/NFAT-Luc reporter cells were incubated with serially diluted anti-CD20 mAbs in the presence of Raji cells. FcγR activation was evaluated by assessing the luminescence intensity.

    Journal: mAbs

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies

    doi: 10.1080/19420862.2019.1608143

    Figure Lengend Snippet: FcγR activation properties of glycoengineered anti-CD20 mAbs. FcγRIIa (a) and FcγRIIIa (b) activation properties of glycoengineered mAbs. Jurkat/FcγRIIa/NFAT-Luc or Jurkat/FcγRIIIa/NFAT-Luc reporter cells were incubated with serially diluted anti-CD20 mAbs in the presence of Raji cells. FcγR activation was evaluated by assessing the luminescence intensity.

    Article Snippet: The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously.

    Techniques: Activation Assay, Incubation

    CDC and C1q-binding activities of glycoengineered anti-CD20 mAbs. (a, b) CDC activities of glycoengineered anti-CD20 mAbs. Raji cells were incubated with 16% human serum and were serially diluted with glycoengineered anti-CD20 mAbs. (a) Percentages of cell lysis plotted against mAb concentrations. (b) CDC activity of 1 µg/ml of G1aF mAb and G1bF mAb mixtures in different ratios. (c) C1q binding of glycoengineered anti-CD20 mAbs. Raji cells were opsonized with anti-CD20 mAbs and incubated with human serum. The cells were stained with FITC-conjugated anti-C1q antibody and the C1q-binding level was analyzed by flow cytometer. Data are presented as mean ± SD ( n = 3).

    Journal: mAbs

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies

    doi: 10.1080/19420862.2019.1608143

    Figure Lengend Snippet: CDC and C1q-binding activities of glycoengineered anti-CD20 mAbs. (a, b) CDC activities of glycoengineered anti-CD20 mAbs. Raji cells were incubated with 16% human serum and were serially diluted with glycoengineered anti-CD20 mAbs. (a) Percentages of cell lysis plotted against mAb concentrations. (b) CDC activity of 1 µg/ml of G1aF mAb and G1bF mAb mixtures in different ratios. (c) C1q binding of glycoengineered anti-CD20 mAbs. Raji cells were opsonized with anti-CD20 mAbs and incubated with human serum. The cells were stained with FITC-conjugated anti-C1q antibody and the C1q-binding level was analyzed by flow cytometer. Data are presented as mean ± SD ( n = 3).

    Article Snippet: The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously.

    Techniques: Binding Assay, Incubation, Lysis, Activity Assay, Staining, Flow Cytometry

    Comparison of structural stabilities of the CH2 domain of glycoengineered anti-CD20 mAbs using HDX/MS . Deuterium uptake plots of peptides Phe245–Leu255 (FLFPPKPKDTL) (a), Leu246–Leu255 (LFPPKPKDTL) (b), and Leu246–Met256 (LFPPKPKDTLM) (c) in the H-chain. (d) Physical representations of the crystal structures (PDB 1HZH) of peptides at Phe245–Met256 (magenta ribbons).

    Journal: mAbs

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies

    doi: 10.1080/19420862.2019.1608143

    Figure Lengend Snippet: Comparison of structural stabilities of the CH2 domain of glycoengineered anti-CD20 mAbs using HDX/MS . Deuterium uptake plots of peptides Phe245–Leu255 (FLFPPKPKDTL) (a), Leu246–Leu255 (LFPPKPKDTL) (b), and Leu246–Met256 (LFPPKPKDTLM) (c) in the H-chain. (d) Physical representations of the crystal structures (PDB 1HZH) of peptides at Phe245–Met256 (magenta ribbons).

    Article Snippet: The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously.

    Techniques:

    Glycan profiles of mAbs with homogeneous glycans. Glycan profiles of commercially available anti-CD20 mAb (a) and glycoengineered G0F mAb (b), G1aF mAb (c), G1bF mAb (d), and G2F mAb (e). The glycan profiles were obtained using HPLC analysis of 2-AB-labeled glycans.

    Journal: mAbs

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies

    doi: 10.1080/19420862.2019.1608143

    Figure Lengend Snippet: Glycan profiles of mAbs with homogeneous glycans. Glycan profiles of commercially available anti-CD20 mAb (a) and glycoengineered G0F mAb (b), G1aF mAb (c), G1bF mAb (d), and G2F mAb (e). The glycan profiles were obtained using HPLC analysis of 2-AB-labeled glycans.

    Article Snippet: The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously.

    Techniques: High Performance Liquid Chromatography, Labeling

    Binding of glycoengineered anti-CD20 mAbs to human FcγRs. SPR analysis was used to measure the binding of anti-CD20 mAbs to human FcγRI, FcγRIIa, and FcγRIIIa. Binding sensorgrams corrected by both the surface blank and buffer injection control are represented.

    Journal: mAbs

    Article Title: Effects of terminal galactose residues in mannose α1-6 arm of Fc-glycan on the effector functions of therapeutic monoclonal antibodies

    doi: 10.1080/19420862.2019.1608143

    Figure Lengend Snippet: Binding of glycoengineered anti-CD20 mAbs to human FcγRs. SPR analysis was used to measure the binding of anti-CD20 mAbs to human FcγRI, FcγRIIa, and FcγRIIIa. Binding sensorgrams corrected by both the surface blank and buffer injection control are represented.

    Article Snippet: The binding of glycoengineered anti-CD20 mAbs to recombinant human FcγRs with C-terminal polyhistidine tag (Cat. No. 10256-H08H, 10374-H08H1, 10259-H08H, 10389-H08H1, and 11046-H08H, Sino Biological) was assessed as described previously.

    Techniques: Binding Assay, SPR Assay, Injection

    Expression of anti-CD20 mAb in transgenic silkworms. ( A ) The protein lysates extracted from MSGs of H line or L line transgenic silkworms were separated by SDS-PAGE followed by staining with CBB or by western blotting with an anti-Human IgG(H + L) antibody. The protein lysates extracted form MSGs of transgenic silkworms that harbored only the Ser1-GAL4 construct were used as negative controls. The numbers above the gels or western blots indicate the line number of each transgenic strain. ( B ) The protein lysates extracted form MSGs or cocoons of the H + L line transgenic silkworms were separated by SDS-PAGE under reducing or non-reducing conditions. Gels were stained with CBB or subjected to protein gel blotting using an anti-Human IgG(H + L) antibody. The lysates obtained from the transgenic silkworms harboring only the Ser1-GAL4 construct were used as a negative control.

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: Expression of anti-CD20 mAb in transgenic silkworms. ( A ) The protein lysates extracted from MSGs of H line or L line transgenic silkworms were separated by SDS-PAGE followed by staining with CBB or by western blotting with an anti-Human IgG(H + L) antibody. The protein lysates extracted form MSGs of transgenic silkworms that harbored only the Ser1-GAL4 construct were used as negative controls. The numbers above the gels or western blots indicate the line number of each transgenic strain. ( B ) The protein lysates extracted form MSGs or cocoons of the H + L line transgenic silkworms were separated by SDS-PAGE under reducing or non-reducing conditions. Gels were stained with CBB or subjected to protein gel blotting using an anti-Human IgG(H + L) antibody. The lysates obtained from the transgenic silkworms harboring only the Ser1-GAL4 construct were used as a negative control.

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Expressing, Transgenic Assay, SDS Page, Staining, Western Blot, Construct, Negative Control

    FcγRIIIa activation and antibody-dependent cell-mediated cytotoxicity (ADCC). ( A ) CD20 binding-dependent activation of FcγRIIIa was measured by using Daudi (target) and Jurkat/FcγRIIIa/NFAT-Luc (effector) cells. Fold increases in luciferase activity were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) The levels of ADCC against Daudi cells were measured by using human PBMCs as effector cells. The percentages of cell lysis were plotted against the mAbs concentration (n = 3, bars indicate SEM).

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: FcγRIIIa activation and antibody-dependent cell-mediated cytotoxicity (ADCC). ( A ) CD20 binding-dependent activation of FcγRIIIa was measured by using Daudi (target) and Jurkat/FcγRIIIa/NFAT-Luc (effector) cells. Fold increases in luciferase activity were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) The levels of ADCC against Daudi cells were measured by using human PBMCs as effector cells. The percentages of cell lysis were plotted against the mAbs concentration (n = 3, bars indicate SEM).

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Activation Assay, Binding Assay, Luciferase, Activity Assay, Concentration Assay, Lysis

    CD20-binding properties. ( A ) Daudi cells were incubated with anti-CD20 mAbs and their binding properties were assessed by flow cytometric analysis using Alexa488-conjugated F(ab')2 anti-human IgG Fc. The mean fluorescence intensities were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) Daudi cells were incubated with DyLight 488-labeled MabThera and serially diluted unlabeled mAbs. Percentages of DyLight 488-labeled MabThera binding were calculated by the mean fluorescence intensities and plotted against the unlabeled mAbs concentration (n = 3, bars indicate SEM).

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: CD20-binding properties. ( A ) Daudi cells were incubated with anti-CD20 mAbs and their binding properties were assessed by flow cytometric analysis using Alexa488-conjugated F(ab')2 anti-human IgG Fc. The mean fluorescence intensities were plotted against the mAbs concentration (n = 3, bars indicate SEM). ( B ) Daudi cells were incubated with DyLight 488-labeled MabThera and serially diluted unlabeled mAbs. Percentages of DyLight 488-labeled MabThera binding were calculated by the mean fluorescence intensities and plotted against the unlabeled mAbs concentration (n = 3, bars indicate SEM).

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Binding Assay, Incubation, Fluorescence, Concentration Assay, Labeling

    Complement-dependent cytotoxicity (CDC). Daudi cells were cultured in the presence of human serum (16%) and anti-CD20 mAbs (1, 3 or 10 µg/ml). The percentages of 7-AAD positive-dead cells were calculated by flow cytometric analysis and represented as the mean + SEM (n = 3). **, p

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: Complement-dependent cytotoxicity (CDC). Daudi cells were cultured in the presence of human serum (16%) and anti-CD20 mAbs (1, 3 or 10 µg/ml). The percentages of 7-AAD positive-dead cells were calculated by flow cytometric analysis and represented as the mean + SEM (n = 3). **, p

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Cell Culture

    Structures of the plasmids used to generate transgenic silkworms. Each plasmid has right and left arms of piggyBac and the 3 × P3-fluorescent gene cassette for a screening marker (EYFP, AmCyan, or DsRed2). Plasmids pBac[UAS_anti-CD20 mAb HC/3 × P3-EYFP] and pBac[UAS_anti-CD20 mAb LC/3 × P3-AmCyan] encode the anti-CD20 mAb H chain gene and the anti-CD20 mAb L chain gene, respectively, under the control of a UAS promoter, and contain an BmNPV- derived hr5 enhancer and an A3-Blasticidin cassette. The anti-CD20 H and L genes were fused to the signal peptide sequence of the sericin 1 gene encoded by exon 1 and 18 bp of exon 2. The plasmid pBac[Ser1-GAL4/3 × P3-DsRed] encodes the GAL4 gene under the control of the sericin1 promoter.

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: Structures of the plasmids used to generate transgenic silkworms. Each plasmid has right and left arms of piggyBac and the 3 × P3-fluorescent gene cassette for a screening marker (EYFP, AmCyan, or DsRed2). Plasmids pBac[UAS_anti-CD20 mAb HC/3 × P3-EYFP] and pBac[UAS_anti-CD20 mAb LC/3 × P3-AmCyan] encode the anti-CD20 mAb H chain gene and the anti-CD20 mAb L chain gene, respectively, under the control of a UAS promoter, and contain an BmNPV- derived hr5 enhancer and an A3-Blasticidin cassette. The anti-CD20 H and L genes were fused to the signal peptide sequence of the sericin 1 gene encoded by exon 1 and 18 bp of exon 2. The plasmid pBac[Ser1-GAL4/3 × P3-DsRed] encodes the GAL4 gene under the control of the sericin1 promoter.

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Transgenic Assay, Plasmid Preparation, Marker, Derivative Assay, Sequencing

    Purification of anti-CD20 mAbs from the transgenic silkworms. ( A ) Anti-CD20 mAbs were purified from the lysates of MSGs or cocoons derived from transgenic silkworms by affinity chromatography using a Protein G column. The lysates (original), flow-through fractions from the Protein G column, and eluted fractions were analyzed by non-reducing SDS-PAGE. ( B ) MabThera and anti-CD20 mAbs derived from transgenic silkworms showed the same migration pattern on non-reducing SDS-PAGE.

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: Purification of anti-CD20 mAbs from the transgenic silkworms. ( A ) Anti-CD20 mAbs were purified from the lysates of MSGs or cocoons derived from transgenic silkworms by affinity chromatography using a Protein G column. The lysates (original), flow-through fractions from the Protein G column, and eluted fractions were analyzed by non-reducing SDS-PAGE. ( B ) MabThera and anti-CD20 mAbs derived from transgenic silkworms showed the same migration pattern on non-reducing SDS-PAGE.

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Purification, Transgenic Assay, Derivative Assay, Affinity Chromatography, SDS Page, Migration

    Fc glycosylation analysis of anti-CD20 mAbs. ( A ) Comparison of mass spectra acquired at the elution point of the glycopeptides of EEQYNSTYR, corresponding to 293–301 amino acids (EU numbering) in the heavy chain of anti-CD20 mAbs. ( B ) Percentage distributions of glycoforms were calculated by the relative peak area average values. Symbols: blue square, N-acetylglucosamine; green circle, mannose; yellow circle, galactose; red triangle, fucose.

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: Fc glycosylation analysis of anti-CD20 mAbs. ( A ) Comparison of mass spectra acquired at the elution point of the glycopeptides of EEQYNSTYR, corresponding to 293–301 amino acids (EU numbering) in the heavy chain of anti-CD20 mAbs. ( B ) Percentage distributions of glycoforms were calculated by the relative peak area average values. Symbols: blue square, N-acetylglucosamine; green circle, mannose; yellow circle, galactose; red triangle, fucose.

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques:

    Binding affinity to human Fc receptors. Binding affinities of anti-CD20 mAbs to FcγRI, FcγRIIIa and FcRn were measured by an SPR analysis. ( A ) Binding sensorgrams corrected for both the surface blank and the buffer injection control are represented. Dashed lines indicate the fitting curves generated by 1:1 binding model for FcγRs and bivalent model for FcRn, respectively. ( B ) Dissociation constant (K D ) values are represented as mean + SD (n = 3). *, p

    Journal: mAbs

    Article Title: Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    doi: 10.1080/19420862.2015.1078054

    Figure Lengend Snippet: Binding affinity to human Fc receptors. Binding affinities of anti-CD20 mAbs to FcγRI, FcγRIIIa and FcRn were measured by an SPR analysis. ( A ) Binding sensorgrams corrected for both the surface blank and the buffer injection control are represented. Dashed lines indicate the fitting curves generated by 1:1 binding model for FcγRs and bivalent model for FcRn, respectively. ( B ) Dissociation constant (K D ) values are represented as mean + SD (n = 3). *, p

    Article Snippet: Serially diluted anti-CD20 mAbs were injected into the flow cells and association and dissociation were monitored.

    Techniques: Binding Assay, SPR Assay, Injection, Generated