Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen

Figures

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  Figure 1. Isolation of rTES-26-specific monoclonal antibodies. (A) Polyclonal phage ELISA of rTES-26 antigen during the biopanning cycles (rounds 1–3). (B) Monoclonal phage ELISA analysis of scFv clones from the helminth library against rTES-26 antigen.

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  Figure 2. Summary of gene usage of rTES-26-specific antibodies and comparisons with previously isolated helminth protein antibodies.

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  Figure 3. Monoclonal antibody gene sequence analysis of rTES-26-specific antibodies. Amino acid distribution.

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  Figure 4. Recombinant monoclonal antibody protein verifications of clone 48. (A) SDS-PAGE. (B) Western blot analysis using anti-His HRP-conjugated antibody. (C) Western blot analysis using StrepTactin–HRP.

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  Figure 5. Recombinant monoclonal antibody protein verifications of clone 49. (A) SDS-PAGE. (B) Western blot analysis using anti-His HRP-onjugated antibody. (C) Western blot analysis using StrepTactin–HRP.

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  Figure 6. Binding verification of recombinant monoclonal antibody proteins. (A) Antigen–antibody Western blot analysis of clones 48 and 49. (B) Antigen–antibody ELISA of clones 48 and 49. Both assays were detected using StrepTactin–HRP.

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  Figure 7. Binding verification of recombinant monoclonal antibody proteins. (A) Native antigen–antibody Western blot analysis of clones 48 and 49. (B) Native antigen–antibody ELISA of clones 48 and 49. Both assays were detected using StrepTactin–HRP and rabbit anti-rTES-26 polyclonal antibody was used as the positive control.

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  Figure 8. Titration ELISA of antibody clones 48 and 49.

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  Figure 9. Specificity ELISA of antibody clones 48 and 49 using other helminth antigens.

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  Figure 10. Sensorgrams of different concentrations (1000 nM to 62.5 nM with 2-fold serial dilution) of the two recombinant monoclonal antibodies. (A) Clone 48. (B) Clone 49. (C) Summary of the binding kinetics of clones 48 and 49 against rTES-26 antigen.

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  Figure 11. Preliminary antigen detection ELISA using human serum samples. Samples 1 and 2 are pooled serum samples from Toxocara spp.-seropositive individuals, and Healthy 1 and 2 are pooled serum samples from healthy individuals.