Immunogenicity of recombinant Lugurus zone pellucida-3 expressed in E.coli and efficacy of its antiserum to affect in vitro mouse sperm-egg binding in vitro / 中国免疫学杂志

ABSTRACT

Objective:Present study was aimed to analyze the immunogenicity of recombinant Lugurus zona pellucida-3 protein (r-LZP3) expressed in E.coli,and to evaluate the efficacy of its antiserum to affect in vitro mouse sperm-egg binding.Methods:Female mouse were immunized using LZP3 DNA vaccine for priming,and boosted with r-LZP3 protein.The antibody response level of LZP3 antisera was determined by ELISA.The immunoreactivity and specificity of the anti-LZP3 antisera were tested by immunoblot, indirect immunofluorescence and immunohistochemistry using native mouse ZP.A sperm-zone pellucida binding assay was used to evaluate the efficacy of anti-LZP3 antisera to inhibit mouse sperm-egg binding in vitro.Results: The result of ELISA showed that LZP3 DNA vaccine was able to induce higher antibody titers in mouse.Antiserum could specifically recognize or bind to r-LZP3 protein expressed in E.coli and native mouse ZP in vitro.The LZP3 antisera and r-LZP3 protein also inhibited sperm-egg binding in vitro.CondusionThese results show that r-LZP3 protein is of strong immunogenicity.