Protein purification using solid-phase extraction on temperature-responsive hydrogel-modified silica beads

Journal of Chromatography. a
Kohei OkuboHideko Kanazawa

Abstract

Recently, the importance of biopharmaceuticals in medical treatments has been increasing, and effective protein purification methods are strongly required for their production. In the present study, a temperature-responsive solid-phase extraction (SPE) column was developed for the purification of proteins without affecting their bioactivity. A temperature-responsive polymer hydrogel-modified stationary phase was prepared by coating aminopropyl silica beads (average diameter, 40-64 μm) with poly(N-isopropylacrylamide) (PNIPAAm)-based thermoresponsive hydrogels. n-Butyl methacrylate and acrylic acid were copolymerized with PNIPAAm as hydrophobic and anionic monomers, respectively. Using these temperature-responsive SPE columns, targeted proteins were retained on the thermoresponsive hydrogel at 40 °C through hydrophobic and electrostatic interactions. After the temperature was reduced from 40 °C to 4 °C, the retained proteins were successfully eluted from the column. Using the temperature-responsive SPE system, lysozyme was successfully separated from ovalbumin without any loss in bioactivity (99.7 ± 0.1%). Rituximab, a monoclonal antibody, was also purified from BSA or hybridoma cell culture medium using the prepared SPE column....Continue Reading

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Citations

Sep 18, 2020·Advances in Colloid and Interface Science·Shixiang LiuHailin Cong
Nov 10, 2020·Analytica Chimica Acta·Kenichi Nagase, Hideko Kanazawa
Feb 2, 2021·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Yutaro MaekawaHideko Kanazawa
Sep 11, 2021·Langmuir : the ACS Journal of Surfaces and Colloids·Patrick L Fosso TeneJürgen Rühe
Sep 3, 2021·Journal of Separation Science·Yutaka Matsuda

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