Methods for screening antibodies for use as immunotoxins

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 29, 435 724, 436503, 436512, 436519, 436547, 436548, 436813, G01N 33563, G01N 33567, G01N 33577

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active

050454510

ABSTRACT:
The present disclosure described an assay for screening monoclonal antibodies for their potential as highly cytotoxic immunotoxins. The assay involves treating cells with dilutions of the test antibody followed by a Fab fragment of a secondary antibody coupled to an A chain toxin ("indirect assay"). The cytotoxicity of the indirect assay is compared to that of the direct assay where the monoclonal antibody is coupled to an A chain toxin. 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.

REFERENCES:
patent: 4360592 (1982-11-01), Weltman
patent: 4664911 (1987-05-01), Uhr et al.
patent: 4689311 (1987-08-01), Weltman
Raso et al., J. Immunol. 125(6): 2610-2616 (1980), "Specific Cytotoxicity of a Human Immunoglobulin-Directed Fab'-Ricin A Chain Conjugate".
Fulton et al., J. Immunol. 136(8): 3103-3109 (1986, "The Effect of Antibody Valency and Lysosomotropic Amines on the Synergy Between Ricin A Chain and Ricin B Chain-Containing Immunotoxins".
Weltman et al., (1987), Can. Res., 47:5552-5556.
Vitetta et al., (1987), Science, 238:1098-1104.
Till et al., (1988), Cancer Res., 48:1119-1123.
Weltman et al., (1986), BioTechniques, 4:224-228.

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