Production and characterization of a human antisperm monoclonal antibody against CD52g for topical contraception in women

Background: Approximately 40% of human pregnancies are unintended, indicating a need for more accept-able effective contraception methods. New antibody production systems make it possible to manufacture reagent-grade human monoclonal antibodies (mAbs) for clinical use. We used the Nicotiana platform to pro-duce a human antisperm mAb and tested its efficacy for on-demand topical contraception. Methods: Heavy and light chain variable region DNA sequences of a human IgM antisperm antibody derived from an infertile woman were inserted with human IgG(1) constant region sequences into an agrobacterium and transfected into Nicotiana benthamiana. The product, an IgG(1) mAb ["Human Contraception Antibody" (HCA)], was purified on Protein A columns, and QC was performed using the LabChip GXII Touch protein characterization system and SEC-HPLC. HCA was tested for antigen specificity by immunofluorescence and western blot assays, antisperm activity by sperm agglutination and complement dependent sperm immobili-zation assays, and safety in a human vaginal tissue (EpiVaginal (TM)) model. Findings: HCA was obtained at concentrations ranging from 0.4 to 4 mg/ml and consisted of > 90% IgG monomers. The mAb specifically reacted with a glycan epitope on CD52g, a glycoprotein produced in the male reproductive tract and found in abundance on sperm. HCA potently agglutinated sperm under a variety of relevant physiological conditions at concentrations >= 6.25 mu g/ml, and mediated complement-dependent sperm immobilization at con-centrations >= 1 mu g/ml. HCA and its immune complexes did not induce inflammation in EpiVaginalTM tissue. Interpretation: HCA, an IgG1 mAb with potent sperm agglutination and immobilization activity and a good safety profile, is a promising candidate for female contraception. (C) 2021 The Author(s). Published by Elsevier B.V.