Date of Award

Summer 1999

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

Program/Concentration

Chemistry

Committee Director

James H. Yuan

Committee Director

Roy L. Williams

Committee Member

Mark S. Elliot

Call Number for Print

Special Collections LD4331.B43 H83

Abstract

Human chorionic gonadotropin (hCG) is well known for early immunological and bioassay tests for early detection of pregnancy. Since the hormone specific beta subunit confers unique biological and immunological specificity to hCG, immunochemical measurement of hCG has achieved greater specificity and sensitivity in recent years by using either the beta subunit as an immunogen or a partially reduced beta subunit polypeptide chain. However, antiserum raised against the entire hCG beta subunit usually cross-reacts to some extent with human luteinizing hormone (hLH). hCG has been found to be present in the extract of tumors and minute quantities of hCG are synthesized by nonpregnant individuals. Therefore, a more specific and sensitive measurement of the hCG beta subunit is expected to improve the early detection of pregnancy and diagnosis of trophoblastic and nontrophoblastic malignancies.

A synthetic peptide of beta hCG residues 109-119 was determined to be a feasible as an antigen due to the linearity of the peptide in native hCG. The structure of the peptide was characterized utilizing molecular modeling techniques. Two anti-hCG hybridoma clones, hCG-N-C2 and hCG-N-Cs, were obtained by a hybridoma method. A beta hCG detecting ELISA system was set up based on a sandwich assay by using specific beta hCG monoclonal antibodies. Specificity of the hCG-N-C2 and hCG-N-C5 was tested in the application of Immulon II microwells. Both antibodies gave positive results on reaction with hCG beta subunit and hCG whole molecule. hCG-N-C2 showed no crossreactivity with hLH while hCG-N-C5 had 13 .7% cross-reactivity with hLH. Thus, the antibodies generated in this research are both specific and sensitive.

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DOI

10.25777/t4sd-j123

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