Metal chelate affinity precipitation: Purification of BSA using poly(N-vinylcaprolactam-co-methacrylic acid) copolymers

This investigation involves the metal chelate affinity precipitation of bovine serum albumin (BSA) using a copper ion loaded thermo-sensitive copolymer. The copolymer of N-vinylcaprolactam with methacrylic acid PNVCL-co-MAA was synthesized by free radical polymerization in aqueous solution, and Cu(II) ions were attached to provide affinity properties for BSA. A maximum loading of 48.1 mg Cu2+ per gram of polymer was attained. The influence of pH, temperature, BSA and NaCl concentrations on BSA precipitation and of pH, ethylenediaminetetraacetic acid (EDTA) and NaCl concentrations on elution were systematically probed. The optimum conditions for BSA precipitation occurred when pH, temperature and BSA concentration were 6.0, 10 °C and 1.0 mg/ml, respectively and the most favorable elution conditions were at pH 4.0, with 0.2 M NaCl and 0.06 M EDTA. The maximum amounts of BSA precipitation and elution were 37.5 and 33.7 mg BSA/g polymer, respectively. It proved possible to perform multiple precipitation/elution cycles with a minimal loss of polymer efficacy. The results show that PNVCL-co-MAA is a suitable matrix for the purification of target proteins from unfractionated materials.

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► Cu(II) was attached to provide affinity properties to thermo-sensitive polymer.
► Cation-loaded copolymer was employed as affinity macroligand.
► Thermo-sensitive macroligand was used for adsorption and elution of protein.
► Precipitation was induced by temperature after protein adsorption.