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Clinical Chemistry 19: 1339-1344, 1973;
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Clinical Chemistry, Vol 19, 1339-1344, Copyright © 1973 by the American Association for Clinical Chemistry

Gel Entrapment of Antibody: A New Strategy for Facilitating Both Manual and Automated Radioimmunoassay

Stuart J. Updike 1, John D. Simmons 1, Douglas H. Grant 1, Judith A. Magnuson 1, and Theodore L. Goodfriend 1

1 Department of Medicine, University of Wisconsin, Center for Health Sciences, Madison, Wis. 53706.

A technique of radioimmunoassay is presented that eliminates pipetting and centrifugation, and excludes interferences by high-molecular-weight materials from the incubation and separation steps. A solid-phase binding reagent is prepared by first entrapping antibody in polyacrylamide gel. This gel is then fragmented, sieved, dried with ethanol or lyophilized, and placed in miniature disposable chromatographic columns. Application of the sample to the intra-gel column compartment is determined by the water regain of the gel. This pipetless method of sample application depends on reproducible aliquots of dry gel particles in every column. A method for preloading radiolabeled hormone and standard hormone into the column is also described. This technique has been successfully applied to the assay of angiotensin I and insulin. Dry antibody—gel stored at room temperature for 26 months has not shown loss of binding activity.


Key Words: angiotensin I • insulin • RIA of low-molecular-weight haptens

Submitted on July 23, 1973






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Copyright © 1973 by the American Association for Clinical Chemistry.