文献类型：期刊文献

英文题名：Recyclable aqueous two-phase system based on two pH-responsive copolymers and its application to porcine circovirus type 2 Cap protein purification

作者：He, Juan[1];Wan, Junfen[1];Yang, Ting[1];Cao, Xuejun[1];Yang, Lin[2]

机构：[1]East China Univ Sci & Technol, Dept Bioengn, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Topfond Pharmaceut Co Ltd, 1 Guangming Rd, Zhumadian 463000, Henan, Peoples R China

年份：2018

卷号：1555

起止页码：113

外文期刊名：JOURNAL OF CHROMATOGRAPHY A

收录：;EI(收录号：20181805129503);WOS:【SCI-EXPANDED(收录号:WOS:000432762900013)】；

基金：The authors sincerely appreciate financial support from National Natural Science Foundation of China (No. 21376078).

语种：英文

外文关键词：Recyclable aqueous two-phase systems; Porcine circovirus type 2 Cap protein; pH-Responsive copolymer; Phase separation mechanism

摘要：Aqueous two-phase system (ATPS) has great potential in industrial applications of bio-separations and bio-reactions. However, its large-scale application is limited by recovery difficulty of phase systems. In this paper, a recyclable ATPS was prepared by two pH-responsive copolymers (P-ADB4.99 and P-MDM7.08) and applied for purification of porcine circovirus type 2 Cap protein fermentation broth (PCV2 Cap protein). Phase separation mechanism was studied by using low-field nuclear magnetic resonance (LF-NMR). The results showed that relatively desirable ATPS were formed successfully when two copolymer concentrations were 4%-6% (w/w) with pH 7.5-8.6. The main parameters, such as copolymer concentration, temperature, pH, type and concentration of salts, were investigated. The results demonstrated that the best ATPS consisted of 4% (w/w) P-ADB4.99 and 6% (w/w) P-MDM7.08, the optimal partition coefficient (K) and extraction recovery (ER) of PCV2 cap protein were 0.25 and 94.2% in the presence of 50 mM Li2SO4,3.53 and 93.4% in the presence of 40 mM KCl, respectively, at 30 degrees C, pH 8.1. The maximum purity of PCV2 Cap protein was 88.4%. In addition, the two phase-forming copolymers could be recycled by adjusting solution pH to their isoelectric points (pI), with recoveries of over 97.5%. (C) 2018 Elsevier B.V. All rights reserved.