Human sperm acrosome membrane-associated protein 1 (hSAMP32) plays an important role in the acrosome reaction, sperm-egg primary binding, secondary binding and fusion processes. However, its spatial structural and invivo antifertility function remain unknown. In this study, we first analysed the physical and chemical characteristics and antigenic epitopes of immunised mice using bioinformatics. Then, we constructed the prokaryotic expression vector pcDNA3.1-hSAMP32 to immunise BALB/c mice invivo. IgG antibodies in the serum were detected, and the litter size of female mice and the number of the hamster eggs penetrated were counted. hSAMP32 was found to contain six hydrophilic regions and a signal peptide beginning at amino acid position 29. The transmembrane region of hSAMP32 was located within amino acids 217-239 with α-helices and random coil structures. We predicted five antigenic epitopes. The molecular weight of hSAMP32 was 59 kDa. Moreover, the results of invivo studies revealed that 56 days after the first immunisation, the litter size was significantly smaller for female pcDNA-3.1(+)-hSAMP32-immunised (mean±s.d. 4.33±1.21) than control mice (9.50±0.55), indicating that the immunocontraception vaccine had an antifertility effect. This experiment presents a theoretical and experimental basis for in-depth study of the hSAMP32 mechanism within the sperm-egg fusing process and for the screening of antigenic epitopes with immunocontraceptive properties.