Department of Chemistry Michigan State University East Lansing, Michigan 48824, USA.
Biomacromolecules. 2010 Apr 12;11(4):1019-26. doi: 10.1021/bm9014792.
Porous nylon membranes modified with poly(acid) brushes and their derivatives can rapidly purify proteins via ion-exchange and metal-ion affinity interactions. Membranes containing poly(2-(methacryloyloxy)ethyl succinate) (poly(MES)) brushes bind 118 +/- 8 mg of lysozyme per cm(3) of membrane and facilitate purification of lysozyme from chicken egg white. Moreover, functionalization of the poly(MES) brushes with nitrilotriacetate (NTA)-Ni(2+) complexes yields membranes that bind poly(histidine)-tagged (His-tagged) ubiquitin with a capacity of 85 +/- 2 mg of protein per cm(3) of membrane. Most importantly, the membranes modified with poly(MES)-NTA-Ni(2+) allow isolation of His-tagged cellular retinaldehyde-binding protein directly from a cell extract in <10 min, and the protein purity is comparable to that achieved with commercial affinity columns. Therefore, porous nylon membranes containing functionalized poly(MES) brushes are attractive candidates for rapid, high-capacity purification of His-tagged proteins from cell extracts.
经聚（酸）刷修饰的多孔尼龙膜及其衍生物可通过离子交换和金属离子亲和相互作用快速纯化蛋白质。含有聚（琥珀酸亚乙酯）（poly（MES））刷的膜可结合每立方厘米膜 118±8mg 的溶菌酶，并促进从鸡蛋白中纯化溶菌酶。此外，聚（MES）刷的功能化与亚氨基二乙酸（NTA）-Ni（2+）配合物生成的膜结合每立方厘米膜 85±2mg 的聚组氨酸标记（His-tagged）泛素的能力。最重要的是，用聚（MES）-NTA-Ni（2+）修饰的膜允许在<10min 内直接从细胞提取物中分离 His 标记的细胞视黄醛结合蛋白，并且蛋白质纯度与商业亲和柱相当。因此，含有功能化聚（MES）刷的多孔尼龙膜是从细胞提取物中快速、高容量纯化 His 标记蛋白的有吸引力的候选物。
