Affinity of human thrombin for insoluble polystyrene substituted with sulfonate,n-hydroxyethylsulfamoyl and n-substituted amidine functions |
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Authors: | S. Khamlichi X. J. Yu D. Muller J. Jozefonvicz S. Roeland R. Fuks |
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Abstract: | A new polymeric material was prepared from polystyrene beads with N-substituted amidines and rendered hydrophilic with sulfonate and N-hydroxyethylsulfamoyl groups. It was synthesized by reacting the p-chlorosulfonyl polystyrene with ethanolamine followed by the Michael-like addition of the terminal OH of the polymer (PSEa) to N-t-butyl-N'-phenylpropenamidine 1 thus leading to the final polymer PSEaAm. Its affinity constant for human α-thrombin calculated from the Langmuir adsorption isotherms was about 106 M−1, a value comparable to the one estimated for polystyrene modified with L-arginine methyl ester. This new material was used as stationary phase in affinity chromatography of thrombin confirming a strong and specific interaction between the positively charged amidinium groups and the enzyme at pH = 7.4 and desorption at pH = 10 and high ionic strength. |
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