Polyclonal anti-chymotrypsin antibodies suitable for oriented immobilization of chymotrypsin

Summary Polyclonal antibodies obtained from the serum of pig immunized by DIP-chymotrypsin were separated into two fractions, anti-chymotrypsin IgG I and anti-chymotrypsin IgG II, by the use of chromatography on biospecific adsorbents prepared by chymotrypsin (CHT) immobilization in different ways. IgG I, which did not decrease the proteolytic activity of CHT, was obtained by biospecific affinity chromatography on a column of Sepharose with CHT attached through an immobilized polyvalent inhibitor, antilysin (AL). IgG II was isolated from the fraction unretarded on the column of CHT-AL-Sepharose by chromatography on a column with CHT directly attached to AH-Sepharose activated by glutaraldehyde. IgG II strongly decreased the proteolytic activity of CHT. Comparison of the proteolytic activity of CHT covalently bound to AH-Sepharose with that of CHT noncovalently intercepted by biospecific sorption to Sepharose with attached anti-CHT-IgG I showed a great advantage of the immobilization of CHT by oriented adsorption.