Biospecific complex formation as a tool for oriented immobilization

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Abstract
The formation of biospecific complexes between the carbohydrate moieties localized on the surface of enzyme molecules and lectins (Hsiao and Royer, Ref.3) or between the appropriate antigenic portions of peptide chains and antibodies (Solomon et al., Ref. 18) permits oriented immobilization to be carried out which makes the catalytic site of the immobilized enzyme perfectly accessible. We have performed such oriented immobilization with carboxypeptidase Y which was attached to concanavalin A ‐ Spheron. On the basis of the results of biospecific affinity chromatography of a conjugate of catalase and dextran carried out by Marshall and Humphreys (Ref. 14) on concanavalin A ‐ Sepharose we prepared active conjugate of chymotrypsin with various dextrans for our investigation of immobilization of chymotrypsin through a bound polysaccharide.The experiments carried out by Solomon et al. (Ref. 18) with the immobilization of carboxypeptidase A via a complex of the enzyme with a suitable monoclonal antibody against carboxypeptidase A showed that the immobilized enzyme had the kinetic properties of native carboxypeptidase A. was more stable and could be used several times. Anti‐chymotrypsin immunoglobulin G(IgG)1 suitable for analogous oriented immobilization of chymotrypsin was isolated from a polyclonal pig antibody against diiso‐propylphosphoryl‐(DIP)‐chymotrypsin by biospecific affinity chromatography on a Sepharose column to which chymotrypsin had been attached through a covalent bond between the surface part of the molecule around the active site and immobilized antilysine, a naturally occurring polyvalent trypsin inhibitor.