The pressing need for the development of new contraceptives targeted to sperm-producing individuals and/or that block sperm function requires new concerted efforts in the development of critical tool compounds to allow researchers to explore reproductive targets without experimental limitations imposed by the absence of critical reagents. The main objective of this proposal is to develop a novel pipeline based on the YCharOS (“Antibody Characterization based on Open Science”) antibody characterization model to focus on antibody reagents of interest for the male contraception research community. Named YCharOS-Repro, this program will prioritize target proteins relevant for non-hormonal contraception (NHC) targeting male-specific proteins. We will establish a dual pipeline for antibody assessment relevant to male contraception. Firstly, we will identify a set of strong candidate proteins to target, prioritizing those for which several commercial antibodies are available. One arm of the proposed pipeline will utilize human cell lines in which expression of the target gene has been confirmed and on which we can then apply Cas9-mediated genome editing to knock out the gene as a negative control. Alternatively, we can use mouse knockout models as our negative control, assuming sufficient protein sequence identity. The second arm will be utilized by using primary reproductive tissues of human and mouse males to test antibody reactivity for both species using the battery of tests optimized for our pipeline (such as Western Blo, Immunofluorescence, and Immunoprecipitation). We will focus specifically on whole testis, isolated germ cell sub-types, and mature epididymal or ejaculated spermatozoa.

The antibody characterization results will be publicly accessible as reports for each target protein through the ZENODO preprint server, followed by appropriate peer-reviewed publications.