scholarly journals High expression of human chorionic gonadotrophin beta-subunit using a synthetic vaccinia virus promoter

2001 ◽  
Vol 26 (3) ◽  
pp. 281-287 ◽  
Author(s):  
A Gupta ◽  
S Chandrasekhar ◽  
R Pal ◽  
S Ahlawat ◽  
O Singh
Keyword(s):  
Vaccinia Virus ◽  
Recombinant Vaccinia Virus ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Beta Subunit ◽  
Synthetic Promoter ◽  
Contraceptive Vaccine ◽  
Virus Rna ◽  
Early Late ◽  
Virus Promoter

We have constructed a recombinant vaccinia virus to express the beta-subunit of human chorionic gonadotrophin (betahCG), a secretory glycoprotein that is used as an antigen for a contraceptive vaccine. The cDNA encoding the subunit was cloned under the control of a synthetic promoter that could be recognised by a vaccinia virus RNA polymerase to direct transcription. The peak expression level of betahCG directed by a late synthetic promoter (Psyn) was 11.5 microg/ml, a level that was at least sixfold higher than that directed by the p7.5 early/late promoter. The expressed protein was correctly processed post-translationally such that it attained a conformation with correctly folded discontinuous epitope(s) similar to that seen in native betahCG.

IMPROVED DIAGNOSIS OF PREGNANCY-RELATED GYNAECOLOGICAL EMERGENCIES BY RAPID HUMAN CHORIONIC GONADOTROPHIN BETA-SUBUNIT ASSAY

1981 ◽  
Vol 88 (2) ◽  
pp. 138-140 ◽  
Author(s):  
M. Seppala ◽  
T. Ranta ◽  
E.-M. Rutanen ◽  
U.-H. Stenman ◽  
K. Tontti ◽  
...  
Keyword(s):  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Beta Subunit

scholarly journals Identification of a heterodimer-specific epitope present in human chorionic gonadotrophin (hCG) using a monoclonal antibody that can distinguish between hCG and human LH

2005 ◽  
Vol 34 (3) ◽  
pp. 879-887 ◽  
Author(s):  
R A Gadkari ◽  
S Roy ◽  
N Rekha ◽  
N Srinivasan ◽  
R R Dighe
Keyword(s):  
Monoclonal Antibody ◽  
Variable Region ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Β Subunit ◽  
Single Chain ◽  
Contraceptive Vaccine ◽  
Mapping Analysis ◽  
Shared Epitopes ◽  
Candidate Antigen

Human chorionic gonadotrophin (hCG) is secreted during early pregnancy and is required for implantation and maintenance of the pregnancy. Active or passive immunoneutralization of hCG results in termination of pregnancy and this forms the basis of the hCG-based female contraceptive vaccine. However, the β subunit of hCG possesses 85% sequence homology with the first 114 amino acids of the β subunit of pituitary human LH (hLH), which is required for ovulation and maintenance of the corpus luteum function during the menstrual cycle. Immunization against hCG or its β subunit leads to generation of antibodies that can neutralize hLH due to many shared epitopes and hence may cause abnormal menstrual cycles. Therefore, it is essential to identify epitopes that are different in the two hormones. In the present study, we report a monoclonal antibody (MAb) specific for hCG that shows no binding to the isolated subunits. Interestingly, the MAb also does not bind hLH at all. The epitope mapping analysis revealed that this antibody recognizes a unique discontinuous epitope present only in the heterodimeric hCG and is distinct from the unique C-terminal extension of hCGβ that is absent in hLHβ. The MAb, either as IgG or its recombinant single-chain variable region fragment, inhibited the response to hCG, but not to hLH. Thus, the epitope recognized by this MAb is an ideal candidate antigen for immunocontraception.

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