A quantitative ELISA was performed to determine the concentration of antibodies. Anti-human IgG kappa antibodies (Thermo Fisher Scientific) were coated on a 96-well Maxisorp plate (100 ng/well) at 4 °C overnight and incubated with a serially diluted antibody in 0.1% PBA at 37 °C for 1 h. The bound antibody was detected with anti-human IgG (Fc-specific)-HRP antibodies (1:10,000, Thermo) as described previously [24 (link)].
Another quantitative ELISA was performed to determine the concentration of purified recombinant preS1 proteins. Serially diluted GST–preS1 (aa 1–56)–strep was coated on the 96-well plate at 4 °C overnight and incubated with mouse anti-strep tag II antibodies (200 ng/well, IBA). The bound antibodies were detected with anti-mouse IgG (Fc-specific)-HRP antibody (1:10,000, Thermo) as described above.
An indirect ELISA was performed to determine the antigen-binding activity of the antibodies. The preS1 antigen coated on the 96-well plate (30 nM/well) was incubated with a serially diluted antibody. The bound antibodies were detected with anti-human IgG (Fc-specific)-HRP antibodies (1:10,000, Thermo), as described above.
Another quantitative ELISA was performed to determine the concentration of purified recombinant preS1 proteins. Serially diluted GST–preS1 (aa 1–56)–strep was coated on the 96-well plate at 4 °C overnight and incubated with mouse anti-strep tag II antibodies (200 ng/well, IBA). The bound antibodies were detected with anti-mouse IgG (Fc-specific)-HRP antibody (1:10,000, Thermo) as described above.
An indirect ELISA was performed to determine the antigen-binding activity of the antibodies. The preS1 antigen coated on the 96-well plate (30 nM/well) was incubated with a serially diluted antibody. The bound antibodies were detected with anti-human IgG (Fc-specific)-HRP antibodies (1:10,000, Thermo), as described above.