No abstract

Problem Requirement of multiple injections of contraceptive vaccines to achieve infertility is one of the important impediments for their application. In the present study, attempts have been made to reduce the number of injections of contraceptive vaccine. Method of study Fusion protein encompassing C‐terminus fragment of sperm protein Sp17 (aa residues 76‐126) and two copies of gonadotropin‐releasing hormone along with T‐cell epitopes and dilysine linkers (abbreviated as Sp17C‐GnRH2) was expressed in Escherichia coli. Its immunogenicity and contraceptive efficacy have been evaluated in female FVB/J mice using different adjuvants and delivery platforms. Results Immunization of female mice with recombinant Sp17C‐GnRH2 (25 μg/injection/mouse) emulsified with squalene‐arlacel A following two injections schedule led to failure of 88.8% immunized animals to conceive, which was not significantly different from mice immunized with same protein along with alum following three injections schedule. To make single‐dose vaccine, poly d,l‐lactic acid–based microparticles (PLA‐MPs) entrapping Sp17C‐GnRH2 were prepared. Immunization of female mice with a combination of soluble Sp17C‐GnRH2 (12.5 μg/injection/mouse) along with Sp17C‐GnRH2 entrapped in PLA‐MPs (12.5 μg/injection/mouse) in alum showed higher antibody titres and contraceptive efficacy as compared to mice immunized with Sp17C‐GnRH2 entrapped in PLA‐MPs alone in alum. Immunization with recombinant Sp17C‐GnRH2 led to long‐term infertility as second mating (150 days after immunization) of various groups of immunized mice showed similar infertility as observed during first mating. Conclusion Single‐dose immunization with PLA‐MPs entrapping Sp17C‐GnRH2 along with soluble recombinant protein in alum generated long‐lasting infertility in female mice.