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A Modification of the Hemolytic Plaque Assay for Use with Protein Antigens¹

Scripps Clinic and Research Foundation, Department of Experimental Pathology, La Jolla, California

Abstract

A method is presented which allows the in vitro enumeration of cells producing antibody against a variety of protein antigens. The proteins are covalently linked to red blood cells by means of a carbodiimide reagent. The concentrations of protein required for the plaque assay are greater than those required for passive agglutination.

The method is simple and sensitive and the results mimic the kinetics of the response that is seen in in vivo assays of serum antibody. Both direct (probably 19 S) and indirect (probably 7 S) plaque-producing cells are detected.

Footnotes

This is publication no. 230 from the Department of Experimental Pathology, Scripps Clinic and Research Foundation, La Jolla, California. The work was supported in part by United States Public Health Service Grant 7007-02, Atomic Energy Commission Contract AT (04-3)-410 and American Cancer Society Grant E-395.

² Supported by United States Public Health Service Training Grant GM-683.

³ Supported by United States Public Health Service Special Fellowship number 2-F3-CA-23, 938-03.

⁴ Supported by United States Public Health Service Research Career Award 5-K6-GM-6936.

⁵ Supported by a Dernham Fellowship of the California Division, American Cancer Society (no. D-100).

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