Non-Specific Binding of Proteins by Substituted Agaroses
Previous results (1–3) showed that at slightly alkaline pH and at relatively low ionic strength (e.g., 0.01–0.05 M Tris-HCl) a large number of negatively charged but unrelated proteins invariably were strongly bound by agarose (Sepharose 4B) substituted with an n-alkylamine or with 4-phenyl-n-butylamine (PBA).l Un-substituted agarose or agarose treated with CNBr, but without subsequent addition of amine, did not bind these proteins. Positively charged protein species generally showed little affinity for the substituted agaroses. Of the proteins tested, the only exception was α-chymotrypsin which, despite its positive charge, showed strong affinity for the PBA-substituted material. By contrast, strong binding did not occur when the ligand was an nalkylamine.
KeywordsIonic Strength Light Absorbance Salt Gradient Original Protein Present Adsorbent
bovine serum albumin
alkylamine with n straight-chain C-atoms
agarose substituted with Cn
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