Patent ID: 6433078
Filing Date: 2002-08-13
Classification: C12N,Y10S

Abstract:
A reversibly soluble, catalytically active enzyme-polymer conjugate made by a process comprising:(a) contacting a free enzyme having at least one free amino group with a modifying agent, the modifying agent having at least one vinyl double bond and an active acylating group, the active group forming an amide bond between the modifying agent and the free amino group of the enzyme and producing a modified enzyme having a free vinyl double bond; (b) recovering the modified enzyme; (c) contacting the modified enzyme with a soluble monomer having the structure R1R2CCR3R4, wherein R1 is selected from hydrogen, carboxy or phenyl moieties, R2 is a hydrogen moiety; R3 is selected from hydrogen, methyl, carboxy, sulfo, or 2-pyridine moieties and R4 is selected from hydrogen, methyl, methoxy, aminopropyl, N,N-dimethylaminopropyl, N,N-diethylaminopropyl, â€ƒor C(O)NR5R6, wherein R5 is selected from hydrogen, methyl, ethyl, propyl, iso-propyl, or propyl, tetrahydro-pyran-2-yl, 2-methoxy-ethyl, R6 is selected from ethyl, propyl, iso-propyl, cyclo-propyl, tetrahydropyrano-2-yl, 2-methoxy-ethyl, 3-methoxy-propyl, 3-iso-propoxy-propyl or â€”C(CH3)2â€”C(O)â€”NHR7, wherein R7 is selected from methyl, ethyl, iso-propyl, 3-methoxy-propyl or 2,2-diethoxy-ethyl, and wherein the monomer is dissolved in a solution and is selected from a group of monomers which when polymerized contain both hydrophobic and hydrophilic regions; and (d) contacting an initiating agent or agents with the monomer, thereby causing a bond to form between the modified enzyme and the monomer and producing an active enzyme-polymer conjugate that reversibly changes its solubility upon a change in the temperature, salt concentration or pH of the solution.