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 of pancreatic ribonuclease A (RNase) has been investigated. The dodecapeptide SRNLTKDRAKPV corresponding to the labile region 32--43 on RNase was synthesized by the solid-phase method. Antiserum raised against the dodecapeptide-bovine serum albumin conjugate showed good cross-reactivity with the peptide and native RNase. RNase immobilized on Sepharose support precoupled either with the antipeptide immunoglobulin (IgG) or anti-RNase IgG proved to be more resistant to thermal inactivation than the soluble enzyme. Besides, stability against inactivation by trypsin at 55 degrees C was markedly high when enzyme was immobilized on the antipeptide IgG support, as compared to the soluble and other immobilized preparations. These results suggest that matrices bearing antibodies recognizing specific labile regions of enzyme may be useful in selectively improving their stability against specific forms of inactivation.
Younus
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Interdisciplinary Biotechnology Unit, Faculty of Life Sciences, Aligarh Muslim University, India.
Owais
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Rao
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Saleemuddin
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D000818:Animals; D000906:Antibodies / Q000276:immunology; D001665:Binding Sites / Q000276:immunology; D003429:Cross Reactions; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D007074:Immunoglobulin G / Q000276:immunology; D009842:Oligopeptides / Q000276:immunology; D010179:Pancreas / Q000201:enzymology*; D011817:Rabbits; D012259:Ribonuclease, Pancreatic / Q000737:chemistry* / Q000276:immunology; D012685:Sepharose; D013696:Temperature; D014357:Trypsin
D000906:Antibodies; D004800:Enzymes, Immobilized; D007074:Immunoglobulin G; D009842:Oligopeptides; D012685:Sepharose; D012259:Ribonuclease, Pancreatic; D014357:Trypsin