Isolation and dynamic hapten binding properties of 2,4,6-triphenyl- N-(4-hydroxyphenyl)-pyridinium (THP-) specific antibodies from bovine colostrum—I : 7S and 19S anti-THP antibodies. Preparation, characterization and analytical application of immunoadsorption

Abstract

Antibodies against the solvatochromic dye 4-(3-aminophenyl)-2,6-diphenyl- N-(4-hydroxy-phenyl)-pyridinium betain (ATHP ∗) we produced by immunization of pregnants cows with an THP-edestin conjugate. Large amounts of 7S and 19S anti-THP antibodies were isolated from the colostrum of one of the immunized animals. The specific isolation was performed with the aid of an immunoadsorbent prepared by coupling ATHP to bovine γ-globulin, previously cross-linked with glutaraldehyde. Various conditions for antibody binding to, and elution from, the adsorbent were studied and compared. Using the adsorbent a test was developed which enabled quantitative determination of anti-THP antibodies, including the non-precipitating ones, which were found to be present in the colostrum in large quantities. The isolated anti-THP antibodies were fractionated by chromatography on Sephadex G-200 into 7S and 19S antibodies. The 7S anti-THP antibodies proved to belong the type IgG s whereas the 19S anti-THP antibodies were composed of 19S IgG s and IgM.