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ELISA-based analysis for reactivity of anti-PspA hkR36A MAbs with recombinant PspAs representing the 6 PspA clades a
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ELISA-based analysis for reactivity of anti-PspA hkR36A MAbs with recombinant PspAs representing the 6 PspA clades a

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: ELISA-based analysis for reactivity of anti-PspA hkR36A MAbs with recombinant PspAs representing the 6 PspA clades a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant

Surface binding of anti-PspA hkR36A MAbs with strains expressing family 1 PspA a

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Surface binding of anti-PspA hkR36A MAbs with strains expressing family 1 PspA a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Binding Assay, Expressing

Relative efficacy of anti-PspAhkR36A MAbs to protect mice against intravenous challenge. CBA/N mice were injected with purified anti-PspAhkR36A MAb M4F4, P1E11, or L5C8 (A and B), B3D12, B3H8, or L5F10 (C and D) or P2A4 or P2B5 (E and F) intraperitoneally at 5 mg/kg body weight (high dose). The corresponding isotype control MAb (IgG1 IC or IgG2a IC) was included in each set as the negative control. One hour later, mice were challenged with 107 CFU of BG8838 (A, C, and E) or 103 CFU of WU2 (B, D, and F), and mouse survival was recorded. The data for the group given anti-PspAhkR36A MAb were compared with those for the respective isotype control MAb using the log rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, statistically not significant.

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Relative efficacy of anti-PspAhkR36A MAbs to protect mice against intravenous challenge. CBA/N mice were injected with purified anti-PspAhkR36A MAb M4F4, P1E11, or L5C8 (A and B), B3D12, B3H8, or L5F10 (C and D) or P2A4 or P2B5 (E and F) intraperitoneally at 5 mg/kg body weight (high dose). The corresponding isotype control MAb (IgG1 IC or IgG2a IC) was included in each set as the negative control. One hour later, mice were challenged with 107 CFU of BG8838 (A, C, and E) or 103 CFU of WU2 (B, D, and F), and mouse survival was recorded. The data for the group given anti-PspAhkR36A MAb were compared with those for the respective isotype control MAb using the log rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, statistically not significant.

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Injection, Purification, Negative Control

Anti-PspAhkR36A MAbs protect mice against pneumococcal infection even when given at a lower dose. CBA/N mice were injected intraperitoneally with 1.25 mg/kg body weight (low dose) of either anti-PspAhkR36A MAb M4F4, P1E11, or L5F10 (A and B) or P2A4 or P2B5 (C and D). The control group was given the respective isotype control MAb (IgG1 IC or IgG2a IC). Mice were challenged with BG8838 (A and C) or WU2 (B and D) 1 h later, and mouse survival was recorded. For other details, refer to the legend to Fig. 2.

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Anti-PspAhkR36A MAbs protect mice against pneumococcal infection even when given at a lower dose. CBA/N mice were injected intraperitoneally with 1.25 mg/kg body weight (low dose) of either anti-PspAhkR36A MAb M4F4, P1E11, or L5F10 (A and B) or P2A4 or P2B5 (C and D). The control group was given the respective isotype control MAb (IgG1 IC or IgG2a IC). Mice were challenged with BG8838 (A and C) or WU2 (B and D) 1 h later, and mouse survival was recorded. For other details, refer to the legend to Fig. 2.

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Infection, Injection

Flow cytometry-based analysis of complement C3 deposition on the surface of pneumococci in the presence of anti-PspA hkR36A MAbs a

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Flow cytometry-based analysis of complement C3 deposition on the surface of pneumococci in the presence of anti-PspA hkR36A MAbs a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Cytometry, Negative Control, Positive Control

In vivo protective efficacies of anti-PspA hkR36A MAbs correlate with the extent of complement deposition a

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: In vivo protective efficacies of anti-PspA hkR36A MAbs correlate with the extent of complement deposition a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: In Vivo, Staining, Activity Assay