Specific immobilization of an enzyme by monoclonal antibody: immobilization of guinea pig liver transglutaminase.
A simple method for enzyme immobilization using a monoclonal antibody raised against the enzyme was studied, taking guinea pig liver transglutaminase as an example. This method consists of two steps; a conjugation of monoclonal antibodies, which bind with the enzyme without inhibitory effect on the activity, with agarose gel bead supports, and immobilization of enzyme proteins onto the conjugated monoclonal antibodies through the antigen-antibody immunoreaction. The following are the advantages of this method. (i) Activity recovery after the immobilization was very high, (ii) enzyme proteins in crude enzyme preparation could be immobilized specifically, and (iii) immobilized enzymes could be replaced easily by fresh enzyme proteins through a urea-buffer treatment.
D000217:Acyltransferases / Q000302:isolation & purification*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004800:Enzymes, Immobilized / Q000302:isolation & purification*; D006168:Guinea Pigs; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011700:Putrescine / Q000032:analysis; D013205:Staphylococcal Protein A / Q000302:isolation & purification; D011503:Transglutaminases
D000911:Antibodies, Monoclonal; D004800:Enzymes, Immobilized; D013205:Staphylococcal Protein A; D000217:Acyltransferases; D011503:Transglutaminases; D011700:Putrescine