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An Assay That Predicts the Ability of Monoclonal Antibodies to Form Potent Ricin A Chain-containing Immunotoxins¹

Department of Microbiology, Southwestern Medical School, University of Texas Health Science Center at Dallas, Dallas, Texas 75235 [M. T., R. D. M., J. W. U., E. S. V.], and the Drug Targeting Laboratory, Imperial Cancer Research Fund Laboratories, London, WC2A 3PX, United Kingdom [P. E. T.]

In this report, we describe an assay for screening monoclonal antibodies for their cytotoxic potential as ricin A chain-containing immunotoxins. The assay involves treating cells with dilutions of the test antibody followed by a Fab fragment of a secondary antibody coupled to ricin A chain ("indirect assay"). The cytotoxicity of the indirect assay is compared to that of the direct assay where the monoclonal antibody is coupled to ricin A chain. Indirect and direct assays were carried out using 14 antibodies and a panel of 8 human and mouse cell types. The two assays showed virtually 100% correlation. The indirect assay, therefore, predicts the potency of a given monoclonal antibody to make an effective immunotoxin and should be useful in screening monoclonal antibodies for use as immunotoxins.

¹ Supported in part by NIH grants CA-48081, CA-28149, a grant from the Welch Foundation (I-947), and a donation from the Meadows Foundation.

² To whom requests for reprints should be addressed, at Department of Microbiology, University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, TX 75235.

Received 9/ 1/87. Revised 11/25/87. Accepted 12/ 2/87.

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