Date of Award

Summer 1982

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

Program/Concentration

Chemistry

Committee Director

Kenton M. Sanders

Committee Director

Roy L. Williams

Committee Member

Billy T. Upchurch

Committee Member

Thomas O. Sitz

Call Number for Print

Special Collections LD4331.C45C65

Abstract

The purpose of this study was to utilize the concept of antibody selective electrodes to developed a new competitive protein binding assay. This is a new concept and a potentially important analytical technique because it combines the advantages of RIA (sensitivity and selectivity) with the advantages of electrical assays (speed and low cost). A conjugate of PGE2 and dibenzo—18- crown-6 (a cation selective ionophore) was synthesized, and incorporated into a plastic membrane. The ionophore conjugate increased the selectivity of a polyvinyl chloride (PVC) membrane to monovalent cations. Transmembrane potential was altered by anti-PGE2 antisera in a concentration dependent manner, whereas non-immune serum was ineffective. The effect of anti-PGE2 antibodies on membrane potential was reversible. Therefore, this antibody electrode was considered as a possible tool to assay prostaglandins. An effective antibody concentration was chosen and PGE2 was added to the antibody solution. "Free" PGE2 competed with the membrane bound PGE2 and reduced the effect of the antibody as a function of "free" PGE2 concentration. Standard curves were generated that could be used to quantitate unknown quantities of PGE2 in solution. The assay system demonstrated specificity toward prostaglandins of the E series. Mechanisms possibly responsible for the effect of antibodies on membrane potential are discussed.

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DOI

10.25777/213f-8z67

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