Application of sperm-specific antibodies for the separation of sperm from cell mixtures

This study was designed to test the suitability of monoclonal antibodies (mAbs) directed against the testicular isoform of the angiotensin-converting enzyme (tACE), coupled with biomagnetic beads, for the extraction of sperm from mixtures with buccal cells. Nine different mAbs recognizing native and denatured form of tACE were tested for immunohistochemical staining of spermatozoa. Three of these mAbs (1E10, 4E3 and 4E10) showing the best staining were then coupled with magnetic beads, and the efficacy and sensitivity of the biomagnetic separation and extraction of spermatozoa was tested. Although the application was limited to swabs stored in PBS buffer instead of air drying and which contain a sufficient amount of intact sperm with mid-piece and flagellum, we have developed an alternative method for the isolation – or at least enrichment – of sperm from epithelial cell mixtures, which is fast, easy and in contrast to the preferential lysis technique, non-toxic.