Premium
Immunogenicity and Contraceptive Efficacy of Escherichia coli ‐Expressed Recombinant Porcine Zona Pellucida Proteins
Author(s) -
Gupta Neha,
Chakrabarti Kausiki,
Prakash Krishna,
Wadhwa Neerja,
Gupta Tripti,
Gupta Satish K.
Publication year - 2013
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/aji.12095
Subject(s) - zona pellucida , immunogenicity , recombinant dna , zona pellucida glycoprotein , western blot , biology , epitope , antibody , microbiology and biotechnology , immune system , toxoid , immunization , virology , andrology , immunology , oocyte , biochemistry , medicine , embryo , gene
Problem To overcome availability of the purified native zona pellucida ( ZP ) glycoproteins for immunocontraception, porcine ZP 3, and ZP 4 were expressed in E. coli . Method of study Purified recombinant proteins were characterized by SDS ‐ PAGE and W estern blot, and immunogenicity and contraceptive efficacy determined in F v B / J female mice. Results Purified ZP 3, ZP 3 with promiscuous T ‐cell epitope of tetanus toxoid, ZP 4 and ZP 4 incorporating promiscuous T ‐cell epitope of bovine RN ase revealed ~44‐, ~49‐, ~53‐, and ~55‐ kD a bands by SDS‐PAGE and Western blot, respectively. Immunization of female mice with recombinant proteins elicited high antibody titers as well as T ‐cell responses. Immune sera recognized mouse oocyte ZP and also inhibited in vitro fertilization. Immunized mice showed significant decrease in fertility. Recombinant proteins were able to recall memory antibody response in female mice primed with porcine native ZP . Conclusion Availability of recombinant porcine proteins will be useful in the development of contraceptive vaccine.