Development of enzyme-linked immunosorbent assay (ELISA) kit: An important biotechnological tool for medical diagnostics and therapeutics


Chandra K. Mittal, Houston Community College System, Department of Biotechnology, Herichson bioMedix, Inc, Houston, TX 77013, John K. Galiotos, Department of Biotechnology & Chemical Laboratory Technology, Houston Community College System-Northeast College, Houston, TX 77013, and Theresa L. Spain, Medical Laboratory Technology, Houston Community College System-Southeast College, 555 Community College Drive, Houston, TX 77013.
One of us has earlier described the development of a radioimmunoassay (RIA) for cyclic AMP and cyclic GMP in biological fluids and cells. The RIA technique, while widely used for disease detection, presents radioactivity hazard, higher regulatory cost and significant disposal problems. The Enzyme-Linked Immunosobent Assays (ELISA), represent the second-generation immunoassay technique and has been replacing the radioimmunoassay (RIA) as the method of choice. The development of an ELISA kit for the detection of a particular disease or drug involves the characterization of disease or drug-specific antigen-antibody interactions as well as binding of the enzyme-linked antigen or antibody. We will describe the data on RIA techniques for cyclic AMP and cyclic GMP as well as experimental protocols used to generate Quality Assurance (QA) data on diagnostic ELISA kits for peptide hormones, herpes simplex virus and prostate specific antigen (PSA). Data on performance characteristics will relate to specificity, reproducibility and sensitivity of the diagnostic ELISA kits.