Protein purification by chemo-selective precipitation using thermoresponsive polymers
Protein purification by chemo-selective precipitation using thermoresponsive polymers
复制标题
使用热响应聚合物通过化学选择性沉淀纯化蛋白质
DOI：
10.1002/bip.23222
复制
发表时间：
2018-06-01
期刊：
影响因子：
2.9
通讯作者：
Ma, Shanyun
中科院分区：
文献类型：
--
作者：
Wu, Yuanzi;Cai, Zhen;Ma, Shanyun
合成了一种可回收的温度响应性高分子-蛋白质生物偶联物，并将其用于纯化具有游离巯基的蛋白质。在目的蛋白的半胱氨酸残基上修饰二硫键引发剂。聚（N-异丙基丙烯酰胺）表现出较低的临界溶液温度从蛋白质生长。所得的蛋白质-聚合物缀合物被成功地热沉淀并与其他蛋白质分离。该方法被证明与牛血清白蛋白的回收率为76.4%。木瓜蛋白酶的酶活性测试验证了可逆聚合物修饰在极端环境下保护蛋白质而不影响蛋白质的功能性。该研究表明，蛋白质的化学选择性富集和纯化具有良好的潜力。
A recoverable and thermoresponsive polymer-protein bioconjugate is synthesized and employed in the purification of protein with free sulfhydryl groups. Initiator with disulphide was modified on the cysteine residue of the target protein. Poly(N-isopropylacrylamide) exhibiting a lower critical solution temperature was grown from the protein. The resulting protein-polymer conjugate was successfully thermoprecipitated and separated from other proteins. The approach was demonstrated with bovine serum albumin with the recycling yield of 76.4%. Enzyme activity test with papain verified the reversible polymer modification protected protein under extreme environments without affecting the functionality of the protein. This study implies the favorable potential of chemo-selective enriching and purification of proteins.