Generating Anti-TIGIT and CD155 Monoclonal Antibodies for Tumor Immunotherapy

Abstract

Many studies have confirmed that the human poliovirus receptor (PVR; CD155) is related to tumor cell migration, invasion, and thus tumor progression. A PVR receptor binds its ligand T cell Ig and the ITIM domain (TIGIT) to inhibit the function of T and NK cells, thereby allowing tumors to evade immune surveillance. In this study, two IgG1 monoclonal antibodies, anti-CD155 and anti-TIGIT, were expressed by the mammalian transient transfection system, then, antibody-dependent cell-mediated cytotoxicity, antibody-binding affinity, and antitumor efficacy were evaluated subsequently in vitro. In this work, protein A affinity chromatography was used for antibodies' purification. Analysis methods included Western blot, enzyme-linked immunosorbent assay, and flow cytometry. Our data suggested that both the two monoclonal antibodies have a purity of higher than 90%, and bound tightly to the antigen with dissociation constant (K _d) and 50% effective concentrations (EC₅₀) below micromolar range. Most notably, these antibodies promote antitumor activity of immune cells in vitro. Therefore, our study laid down the foundation for subsequent in vivo experiments for further evaluation.

Keywords

monoclonal antibodies - TIGIT - PVR - tumor immunotherapy - antibody-dependent cell-mediated cytotoxicity

Ethics Statement

The separation of PBMCs from healthy donors abides by the relevant agreements of the Changhai Hospital of Shanghai, and it conforms to the provisions of the Declaration of Helsinki in 1995 (16). Informed consent was obtained from all individual participants included in the study.

Publikationsverlauf

Eingereicht: 27. April 2022

Angenommen: 13. Juli 2022

Artikel online veröffentlicht:
02. September 2022

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