Purification of anti-chymotrypsin antibodies for the preparation of a bioaffinity matrix with oriented chymotrypsin as immobilized ligand.
Polyclonal antibodies suitable for the oriented immobilization of chymotrypsin were prepared by chromatography on a bioaffinity matrix which had the enzyme immobilized through its active site to antilysin, covalently linked to bead cellulose. After periodate oxidation of their carbohydrate moieties, the isolated antibodies were coupled to a hydrazide derivative of bead cellulose. The periodate oxidation step, which led to greater efficiency and stability of the immunosorbent, had no deleterious effect on antibody activity as assessed by ELISA. Addition of chymotrypsin to the immunosorbent yielded an enzymically active bioaffinity matrix with the optimum molar enzyme/antibody ratio of 2.
Bílková
Z
Z
Department of Biological and Biochemical Sciences, University of Paradubice, Czech Republic.
Churácek
J
J
Kucerová
Z
Z
Turková
J
J
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D002482:Cellulose; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D002918:Chymotrypsin / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D000939:Epitopes; D007164:Immunosorbents / Q000138:chemical synthesis; D008024:Ligands; D013552:Swine
D000906:Antibodies; D000939:Epitopes; D007164:Immunosorbents; D008024:Ligands; D002482:Cellulose; D002918:Chymotrypsin