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Applied Biochemistry and Biotechnology, Vol.68, No.1-2, 121-133, 1997
Imidazole - A New Ligand for Metal Affinity Precipitation - Precipitation of Kunitz Soybean Trypsin-Inhibitor Using Cu(II)-Loaded Copolymers of 1-Vinylimidazole with N-Vinylcaprolactam or N-Isopropylacrylamide
Kunitz soybean trypsin inhibitor (STI) was specifically coprecipitated during precipitation of Cu(II)-loaded copolymers induced by increase in temperature and ionic strength. The copolymers used consisted of 1-vinylimidazole and N-vinylcaprolactam or N-isopropylacrylamide. The elution of STI was achieved by solubilization of the STI-Cu(II)-polymer complex in the presence of an excess of the competing ligand, imidazole, and a subsequent precipitation of the polymer with STI remaining free in solution in a purified form as judged by Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). To the best of our knowledge this is the first reported successful metal affinity precipitation of protein in a heterobifunctional format.
Keywords:DEPENDENT INTERACTION CHROMATOGRAPHY;PROTEIN SURFACE DOMAINS;IMMUNOGLOBULIN-G;BINDING;HISTIDINE;SILICA;THERMOPRECIPITATION;PURIFICATION;VARIANTS;POLYMER