Revolutionizing sexed sorting sperm using scFv antibodies combined with microbeads for porcine sexed semen.

This study aimed to evaluate porcine sperm sex selection using scFv antibodies combined with magnetic-activated cell sorting (MACS) to optimize X/Y sperm separation while maintaining essential sperm quality parameters, including viability, motility, membrane integrity, and apoptosis, with the goal of developing a novel method for swine sexing technology. The H4L4 scFv antibody, at a concentration of 1 mg/mL, effectively bound to PLA microbeads (HL-beads), achieving separation efficiencies of 75.4 ± 2.30 % for X-sperm and 78.6 ± 1.94 % for Y-sperm. FTIR confirmed the binding of H4L4 scFv to microbead carboxylic acid groups, and SEM verified sperm binding. In quality assessments, the X-enriched fraction (XF) presented a sperm quality comparable to that of conventional semen (CONV) and the negative control (NC). However, the Y-enriched fraction (YF) was lower in quality than the other groups were on Day 3 of storage. Apoptosis analysis revealed no significant difference in the number of necrotic cells among the XF, CONV, and NC groups. The XF group achieved a 76.1 % enriched X-sperm ratio, whereas the YF group achieved a 78.4 % enriched Y-sperm ratio. This method enhances the X-sperm proportion in the XF group with an acceptable quality for farm applications. These results demonstrate the potential of HL-beads for porcine sperm sexing, offering a promising approach for improving sex ratios in swine production.