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Isolation and Validation of Anti-B7-H4 scFvs from an Ovarian Cancer scFv Yeast-Display Library.
Methods In Molecular Biology
B7-H4 (VTCN1, B7x, B7s) is an inhibitory modulator of T-cell response implicated in antigen tolerization. As such, B7-H4 is an immune checkpoint of potential therapeutic interest. To generate anti-B7-H4 targeting reagents, we isolated antibodies by differential cell screening of a yeast-display library of recombinant antibodies (scFvs) derived from ovarian cancer patients and we screened for functional scFvs capable to interfere with B7-H4-mediated inhibition of antitumor responses. We found one antibody binding to B7-H4 that could restore antitumor T cell responses. This chapter gives an overview of the methods we developed to isolate a functional anti-B7-H4 antibody fragment.
Animals, Antigen Presentation, B-Lymphocytes/immunology, Female, Gene Library, Humans, Mice, Ovarian Neoplasms/blood, Ovarian Neoplasms/immunology, Saccharomyces cerevisiae/genetics, Saccharomyces cerevisiae/metabolism, Single-Chain Antibodies/genetics, Single-Chain Antibodies/isolation & purification, T-Lymphocytes/immunology, V-Set Domain-Containing T-Cell Activation Inhibitor 1/metabolism, Xenograft Model Antitumor Assays
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