Journal: Microbiology Spectrum

Article Title: Isolation and Characterization of Human Monoclonal Antibodies to Pneumococcal Capsular Polysaccharide 3

doi: 10.1128/Spectrum.01446-21

Figure Lengend Snippet: HumAb binding to pneumococcal polysaccharide 3 (PPS3) by ELISA. Binding as reflected by absorbance at 405 is shown on the y axis for the humAb concentrations shown on the x axis for each humAb. Results are representative of 3 independent experiments ( n = 2). The numerical 50% effective concentration (EC 50 ) for each humAb is indicated to the right of the panel depicting binding curves of all humAbs.

Article Snippet: Concentrations of fluorescently conjugated PPS3 (PPS3-PE) (Fina BioSolutions) were incubated with ST3 mouse hybridoma cells ( ) with or without unlabeled PPS3 (25 μg/well).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Microbiology Spectrum

Article Title: Isolation and Characterization of Human Monoclonal Antibodies to Pneumococcal Capsular Polysaccharide 3

doi: 10.1128/Spectrum.01446-21

Figure Lengend Snippet: Heavy and light chain VDJ gene usage and CDR3 sequences for all PPS3 humAbs

Article Snippet: Concentrations of fluorescently conjugated PPS3 (PPS3-PE) (Fina BioSolutions) were incubated with ST3 mouse hybridoma cells ( ) with or without unlabeled PPS3 (25 μg/well).

Techniques:

Journal: Microbiology Spectrum

Article Title: Isolation and Characterization of Human Monoclonal Antibodies to Pneumococcal Capsular Polysaccharide 3

doi: 10.1128/Spectrum.01446-21

Figure Lengend Snippet: HumAb binding and agglutination of humAbs with light chain swaps. (A) HumAbs (native) with their LC swaps were generated and tested by ELISA for binding reactivity to purified PPS3 and B2. Absorbance at 405 is shown on the y axis for the humAb concentrations shown on the x axis for each humAb. The 50% effective concentration (EC 50 ) is depicted on the graph. Results are representative of 3 independent experiments ( n = 2). ST3 strain B2 was incubated with increasing concentrations of humAbs (C10, C10 LC swap [C10 H C27 L ] or C27, C27 LC swap [C27 H C10 L ]) or control IgG1 and analyzed by flow cytometry. (B) Representative FACS dot plots showing percent agglutination of the indicated native humAb or LC swap at various concentrations. (C) Line graph depicting percent agglutination on the y axis for concentrations of indicated humAbs and LC swaps on the x axis. Results are representative of 2 independent experiments ( n = 2 per condition). Differences were determined by one-way ANOVA; at 10 μg/ml (C10 versus IgG1, C10 versus C10 LC swap [C10 H C27 L ], C10 versus C27, and C10 versus C27 LC swap [C27 H C10 L ]; ** * , P < 0.001), at 20 μg/ml (C10 versus IgG1, C10 versus C10 LC swap [C10 H C27 L ], C10 versus C27, and C10 versus C27 LC swap [C27 H C10 L ]; * , P < 0.05).

Article Snippet: Concentrations of fluorescently conjugated PPS3 (PPS3-PE) (Fina BioSolutions) were incubated with ST3 mouse hybridoma cells ( ) with or without unlabeled PPS3 (25 μg/well).

Techniques: Binding Assay, Agglutination, Generated, Enzyme-linked Immunosorbent Assay, Purification, Concentration Assay, Incubation, Control, Flow Cytometry

Journal: Microbiology Spectrum

Article Title: Isolation and Characterization of Human Monoclonal Antibodies to Pneumococcal Capsular Polysaccharide 3

doi: 10.1128/Spectrum.01446-21

Figure Lengend Snippet: Summary of in vitro and in vivo functions for all PPS3 humAbs ^a

Article Snippet: Concentrations of fluorescently conjugated PPS3 (PPS3-PE) (Fina BioSolutions) were incubated with ST3 mouse hybridoma cells ( ) with or without unlabeled PPS3 (25 μg/well).

Techniques: In Vitro, In Vivo, Binding Assay, Agglutination, Control