Fig. 3.

Rocket immunoelectrophoresis of MOSPD2 antigen showed reaction against ASA(s) in the seminal plasma of infertile LCS/BS sample.(a) Representative image of Ag-Ab reaction using RIEP. Ag(s) used in this reaction were obtained from sperm of fertile donor (first lane), S. pyogenes (second lane) and synthesized MOSPD2 antigen (third lane). 100 ​μg/ml concentration of sperm and bacteria Ag(s) each were used as positive controls for 100 ​μg/ml MOSPD2 Ag. The seminal plasma obtained from the infertile men with LCS/BS contained ASA(s) for which Ag-Ab reaction was clearly visible in the form of white precipitation peaks or, rockets against the CBB stained agarose gel using RIEP. The presence of ASA(s) against the purified MOSPD2 antigen was validated with the highest rocket peak than the controls. The white line is the length of the rocket and orange line represented the end of the peak on the gel. (b) Representative image of concentration dependent Ag-Ab reaction of synthesized MOSPD2 antigen. Experiment was repeated thrice and height of peaks is average of three independent assays. ASA: Antisperm Antibodies, MOSPD2: Motile Sperm Domain- Containing Protein 2, LCS: Leukocytospermia, BS: Bacteriospermia, Ag: Antigen, CBB: Coomassie Brilliant Blue, RIEP: Rocket Immunoelectrophoresis.