Stabilization of pancreatic ribonuclease A by immobilization on Sepharose-linked antibodies that recognize the labile region of the enzyme

The stabilizing potential of the antibodies recognizing the labile region o f pancreatic ribonuclease A (RNase) has been investigated. The dodecapeptid e SRNLTKDRAKPV corresponding to the labile region 32-43 on RNase was synthe sized by the solid-phase method. Antiserum raised against the dodecapeptide -bovine serum albumin conjugate showed good crossreactivity with the peptid e and native RNase, RNase immobilized on Sepharose support precoupled eithe r with the antipeptide immunoglobulin (IgG) or anti-RNase IgG proved to be more resistant to thermal inactivation than the soluble enzyme. Besides, st ability against inactivation by trypsin at 55 degreesC was markedly high wh en enzyme was immobilized on the antipeptide IgG support, as compared to th e soluble and other immobilized preparations. These results suggest that ma trices bearing antibodies recognizing specific labile regions of enzyme may be useful in selectively improving their stability against specific forms of inactivation. (C) 2001 Elsevier Science B.V. All rights reserved.