Preparation of anti-human TREM2 antibody and preliminary study of its biological activity
Objective To prepare antibodies against human triggering receptor expressed on myeloid cells(TREM2)and evaluate their bioactivity.Methods Monoclonal antibodies against human TREM2 were obtained by hybridoma technique,and the monoclonal antibodies with the best binding effect were selected and humanized to obtain chimeric antibodies.Subsequently,the chimeric antibodies were humanized,and the binding activity of the antibody to antigen were determined by enzyme-linked immunosorbent assay and flow cytometry.The binding kinetics of the humanized antibody to human TREM2 was detected by bio-layer interferometry.The cell-dependent cytotoxicity of the humanized antibody was evaluated with an antibody-dependent cell-mediated cytotoxicity(ADCC)biological activity assay based on the reporter gene.In vivo efficacy experiments were conducted in immunodeficient mice.Results The hybridoma cell lines stably expressing antibodies against TREM2 were successfully screened by hybridoma technology.The further humanized antibody,h7B6-11,had a purity over 95%and a high affinity with TREM2-His protein.The ADCC activity of the antibody h7B6-11 against 293T-TREM2 cells was much stronger than that of the control antibody.The in vivo efficacy experiments showed that the combination of h7B6/11 and programmed cell death protein 1(PD1)antibody greatly inhibited the tumor growth.Conclusion The antibodies possess high binding affinity and excellent bioactivity,which may become prospective therapeutic immunity drugs.
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