Purification of two specific antibodies against drug and carrier protein molecules.

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Abstract
Affinty purification procedures for antibodies specific to a drug and to a carrier protein were examined in detail with the use of three enzyme immunoassays (EIAs). Many blasticidin S (BLS) molecules were coupled to an affinity ligand with pig serum albumin as a spacer protein. The influence of the spacer on the affinity purification of an antibody specific to BLS was quantitatively analyzed. The antibody showed higher binding to BLS bound to the solid matrix with a carrier protein than without. The stability of specific antibody in six representative eluents, used for affinity chromatography of specific antibodies, was examined, and 0.1 M potassium chloride-0.008 N hydrochloric acid buffer was selected as the preferred eluent based on the stability of the specific antibody and convenience in handling. The ability of the buffer to elute the specific antibody from an affinity column was studied, and was improved by modifying the concentration of potassium chloride in the buffer to 0.3 M. Complete purification of anti-BLS antibody was performed by affinity chromatography under the chosen condition. The specific anti-BLS and anti-carrier protein antibodies were purified quantitatively. Formation of denatured specific antibody was hardly detected by a sensitive EIA, under the chosen conditions. The purity of the standard anti-BLS antibody was demonstrated by the affinity chromatographic method with the aid of two EIAs for rabbit immunoglobulin G and for BLS.