Affinity chromatography-based immunoassays
Abstract
Affinity chromatography-based immunoassays were developed in order to meet the analytical requirements of biotechnology. Dual-column immunoassay (DCIA) consisted of an immunoaffinity and a conventional analytical chromatography column, coupled with a switching valve. DCIA was used for performing direct and competitive inhibition of binding immunoassays of single and multicomponent unlabeled antigen samples. DCIA was developed to facilitate the determination of (i) protein concentration in a complex mixture, (ii) the purity of a protein contaminated with variant species of similar structure and (iii) immune response (antibody titer) of an antiserum. Sequential addition competitive binding immunoassays using protein A affinity chromatography were used to determine the concentration of a protein in a complex mixture. Sequential addition immunoassays were very rapid ($<$1 min) and sensitive (0.7 pmol human transferrin).
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Bioengineering|Analytical chemistry
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