An all-aqueous route to polymer brush-modified membranes with remarkable permeabilites and protein capture rates.
An all-aqueous route to polymer brush-modified membranes with remarkable permeabilites and protein capture rates.
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DOI：
10.1016/j.memsci.2011.10.022
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发表时间：
2012-02-01
影响因子：
9.5
通讯作者：
Bruening, Merlin L.
中科院分区：
文献类型：
--
作者：
Anuraj, Nishotha;Bhattacharjee, Somnath;Geiger, James H.;Baker, Gregory L.;Bruening, Merlin L.
微孔膜对蛋白质纯化很有吸引力，因为对流能迅速将蛋白质带到结合位点。然而，这种膜的低结合能力限制了它们的应用。这项工作报告了一种快速的水处理方法，可以制造出结合大量蛋白质的高渗透性聚合物刷修饰膜。该合成方法包括在羟基化尼龙膜上吸附大引发剂10分钟，随后从固定引发剂中进行5分钟的2-（甲基丙烯酰氧基）琥珀酸乙酯的水原子转移自由基聚合以形成聚（酸）刷。与硅烷引发剂聚合相比，这一过程可能会产生更膨胀、密度更低的毛刷，因此需要更少的聚合物来实现相同的结合能力。与固定化硅烷引发剂生长刷法制备的类似膜相比，聚酸膜的渗透性提高了4倍。这些含有刷状的尼龙膜在溶液停留时间短至35 ms的情况下结合120 mg/cm3的溶菌酶，当用三乙酸硝基(NTA)-Ni2+络合物功能化时，它们捕获85 mg/cm3的组氨酸标记（His-tagged）泛素。此外，NTA-Ni2+功能化膜直接从细胞提取物中分离出his标记的肌醇-1-磷酸合成酶，并显示出his标记蛋白的回收率为90%。
Microporous membranes are attractive for protein purification because convection rapidly brings proteins to binding sites. However, the low binding capacity of such membranes limits their applications. This work reports a rapid, aqueous procedure to create highly permeable, polymer brush-modified membranes that bind large amounts of protein. The synthetic method includes a 10-min adsorption of a macroinitiator in a hydroxylated nylon membrane and a subsequent 5-min aqueous atom transfer radical polymerization of 2-(methacryloyloxy)ethyl succinate from the immobilized initiator to form poly(acid) brushes. This procedure likely leads to more swollen, less dense brushes than polymerization from silane initiators, and thus requires less polymer to achieve the same binding capacity. The hydraulic permeability of the poly(acid) membranes is 4-fold higher than that of similar membranes prepared by growing brushes from immobilized silane initiators. These brush-containing nylon membranes bind 120 mg/cm3 of lysozyme using solution residence times as short as 35 ms, and when functionalized with nitrilotriacetate (NTA)-Ni2+ complexes, they capture 85 mg/cm3 of histidine6-tagged (His-tagged) Ubiquitin. Additionally the NTA-Ni2+-functionalized membranes isolate His-tagged myo-inositol-1-phosphate synthase directly from cell extracts and show >90% recovery of His-tagged proteins.
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DOI：
10.1038/nbt0788-779
发表时间：
1988-07-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
BRANDT, S;GOFFE, RA;ZALE, SE
通讯作者：
ZALE, SE
影响因子：
5.5
作者：
Matyjaszewski, K;Miller, PJ;Pakula, T
通讯作者：
Pakula, T
影响因子：
9.5
作者：
Ma, Zuwei;Masaya, Kotaki;Ramakrishna, Seeram
通讯作者：
Ramakrishna, Seeram
影响因子：
5.5
作者：
ITO, Y;INABA, M;IMANISHI, Y
通讯作者：
IMANISHI, Y
影响因子：
5.5
作者：
Jain, Parul;Dai, Jinhua;Bruening, Merlin L.
通讯作者：
Bruening, Merlin L.