Reversible protection of disulfide bonds followed by oxidative folding render recombinant hCGbeta highly immunogenic.
Active immunization of women against human chorionic gonadotropin (hCG) has been considered as a promising option for contraception. However, prototype hCG vaccines based on natural sources of antigen are expected to be costlier for use by common people. In the present report, a functionally active, cost-effective antigen of bacterial origin has been described. Sulfonation of thiol groups of the protein, anion-exchange purification, refolding with concomitant formation of disulfide bonds in the presence of cysteamine-cystamine redox buffer, and slow removal of denaturant resulted in 95% homogeneous, monomeric form of the antigen. The recombinant processed antigen [CGbeta(p)] obtained this way was highly immunopotent. Cellular DNA and endotoxin contaminants were appreciably low in the final product. The immunogenic response was drastically reduced with the unprocessed antigen. This finding envisages better prospect of a cost-effective hCG vaccine for birth control.
Mukhopadhyay
A
A
National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India. amukhopa@notes.mdacc.tmc.edu
D000818:Animals; D000906:Antibodies / Q000097:blood; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D003269:Contraception, Immunologic; D003538:Cystamine / Q000737:chemistry; D003543:Cysteamine / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D004220:Disulfides / Q000737:chemistry* / Q000276:immunology; D004731:Endotoxins / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D000882:Haplorhini; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D010084:Oxidation-Reduction; D011489:Protein Denaturation / Q000276:immunology; D017510:Protein Folding; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D015203:Reproducibility of Results; D020539:Sequence Analysis, Protein; D013447:Sulfites / Q000737:chemistry; D013776:Tetrathionic Acid / Q000737:chemistry
D000906:Antibodies; D018997:Chorionic Gonadotropin, beta Subunit, Human; D004269:DNA, Bacterial; D004220:Disulfides; D004731:Endotoxins; D011994:Recombinant Proteins; D013447:Sulfites; D003543:Cysteamine; D013776:Tetrathionic Acid; D003538:Cystamine; C025026:sodium sulfite