PROBLEM: Monoclonal anti-capacitated sperm antibody has been used as a prob
e to identify, isolate, and characterize specific, fertilization-related an
tigen with some characteristic features that point to its possible signific
ance in immunocontraception.
METHOD OF STUDY: Fast protein liquid chromatography (FPLC), enzyme-linked i
mmunosorbent assay (ELISA), sodium dodecyl sulfate polyacrylamide gel elect
rophoresis (SDS-PAGE), and isoelectrofocusing were used for isolation, immu
nochemical and physicochemical characterization of a monoclonal antibody (m
Ab) 1F10 cognate antigen. Sperm-zone pellucida binding and hemizona assay w
ere used for resting the biological roles of mAb 1F10 and Ag 1F10 in boar a
nd human fertilization processes.
RESULTS: The AE 1F10 was found to be eluted in the eighth protein peak of F
PLC-fractionated Nonidet P40 (NP40) extracts of capacitated boar spermatozo
a. The SDS-PAGE and immunoelectrofocusing experiments showed that Ag 1F10 i
s a protein composed of a single peptide chain with a relative molecular ma
ss of 68/70 kDa and an isoelectric point of 3.5. It was demonstrated that t
he zona binding activity of spermatozoa preincubated in the presence of mAb
1F10 was significantly inhibited both in porcine and human in vitro fertil
ization (IVF) systems. A dose-dependent manner of inhibition of the sperm/l
igand activities of porcine and human zone pellucida was observed when the
effect of purified Ag 1F10 was investigated by its preincubation with zona
CONCLUSIONS: It is assumed that the protein bearing the epitope recognized
by mAb 1F10 may be accepted as one of the molecules with receptor function
in sperm-zone pellucida interaction during fertilization.