Online ISSN: 1884-4111 Print ISSN: 0033-8303
Radioisotopes 67(7): 321-328 (2018)


アクリル酸グラフト繊維を用いた高濃度なリン酸緩衝液中でのリゾチームの高容量吸着Acrylic Acid-grafted Fibers Enable High-capacity Binding of Lysozyme Dissolved in High-concentration Phosphate Buffer

千葉大学工学部Department of Applied Chemistry and Biotechnology, Chiba University ◇ 263–8522 千葉市稲毛区弥生町1–33 ◇ 1–33 Yayoi-cho, Inage-ku, Chiba Pref. 263–8522, Japan

受付日:2018年1月11日Received: January 11, 2018
受理日:2018年3月19日Accepted: March 19, 2018
発行日:2018年7月15日Published: July 15, 2018

高濃度なリン酸緩衝液中でリゾチームを高容量に吸着する弱酸性カチオン交換繊維を作製するために,放射線グラフト重合法を適用して,ナイロン-6繊維へアクリル酸をグラフト重合した。得られた弱酸性カチオン交換繊維をカラムへ充填し,50~250 mMリン酸緩衝液(pH 6.0)に溶かしたリゾチームを空間速度20 h−1で流通し,動的吸着容量(DBC)を測定した。市販の弱酸性カチオン交換ビーズ(CM Sepharose® Fast Flow, GE Healthcare製)充填カラムのDBCは,緩衝液濃度の増大に伴って88から9 mg/mL-bedに減少した。一方,弱酸性カチオン交換繊維充填カラムのDBCは,緩衝液濃度50から200 mMの範囲で一定の値(200 mg/mL-bed)を示した。

To prepare a weakly acidic cation-exchange fiber capable of binding lysozyme dissolved in a high-concentration phosphate buffer, acrylic acid (AA) was graft-polymerized onto a nylon-6 fiber by radiation-induced graft polymerization. The dynamic binding capacity (DBC) of the bed charged with the AA-grafted fiber at a space velocity of 20 h−1 was determined in a flow-through mode for lysozyme solutions buffered with 50–250 mM phosphate solution (pH 6.0). The DBC of the CM-Sepharose-FF-bead-packed column decreased from 88 to 9 mg/mL-bed with increasing buffer concentration. In contrast the AA-grafted-fiber-packed column exhibited a constant value of 200 mg/mL-bed independent of the buffer concentration from 50 to 200 mM.

Key words: radiation-induced graft polymerization; dynamic binding capacity; high-concentration phosphate buffer; weakly acidic cation-exchange fiber; protein purification; acrylic acid-grafted fiber; lysozyme adsorption

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