Herdt Aimee R, Kim Byeong-Su, Taton T Andrew
University of Minnesota, Department of Chemistry, 207 Pleasant Street SE, Minneapolis, Minnesota 55455, USA.
Bioconjug Chem. 2007 Jan-Feb;18(1):183-9. doi: 10.1021/bc060215j.
This paper describes the bioconjugation of histidine-tagged enzymes and other proteins to the surface of composite "magnetomicelles" consisting of magnetic gamma-Fe2O3 nanoparticles encapsulated within cross-linked polystyrene-block-polyacrylate copolymer micelle shells. Free carboxylic acid groups on the magnetomicelle surface were converted to Cu2+-iminodiacetic acid (IDA) for protein capture. The conjugation of T4 DNA ligase and enhanced green fluorescent protein to magnetomicelles revealed that proteins were captured with a high surface density and could be magnetically separated from reaction mixtures and subsequently released from the nanoparticle surface. Additionally, bioconjugation of T7 RNA polymerase yielded a functional enzyme that maintained its biological activity and could be recycled for up to three subsequent transcription reactions. We propose that protein-magnetomicelle bioconjugates are effective for protein bioseparation and enzymatic recycling and further strengthen the idea that nanoparticle surfaces have utility in protein immobilization.
本文描述了组氨酸标记的酶和其他蛋白质与复合“磁性胶束”表面的生物共轭，该复合“磁性胶束”由包裹在交联聚苯乙烯-嵌段-聚丙烯酸酯共聚物胶束壳内的磁性γ-Fe2O3纳米颗粒组成。磁性胶束表面的游离羧酸基团被转化为用于蛋白质捕获的Cu2+-亚氨基二乙酸（IDA）。T4 DNA连接酶和增强型绿色荧光蛋白与磁性胶束的共轭表明，蛋白质以高表面密度被捕获，并且可以从反应混合物中磁性分离出来，随后从纳米颗粒表面释放。此外，T7 RNA聚合酶的生物共轭产生了一种功能性酶，该酶保持其生物活性，并且可以循环用于多达三个后续转录反应。我们提出蛋白质-磁性胶束生物共轭物对于蛋白质生物分离和酶循环是有效的，并进一步强化了纳米颗粒表面在蛋白质固定化方面具有实用性的观点。
