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蛋白质制造仪:一种用于高通量平行纯化的自动化系统。

The Protein Maker: an automated system for high-throughput parallel purification.

作者信息

Smith Eric R, Begley Darren W, Anderson Vanessa, Raymond Amy C, Haffner Taryn E, Robinson John I, Edwards Thomas E, Duncan Natalie, Gerdts Cory J, Mixon Mark B, Nollert Peter, Staker Bart L, Stewart Lance J

机构信息

Seattle Structural Genomics Center for Infectious Disease (http://www.ssgcid.org), USA.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Sep 1;67(Pt 9):1015-21. doi: 10.1107/S1744309111028776. Epub 2011 Aug 13.

Abstract

The Protein Maker is an automated purification system developed by Emerald BioSystems for high-throughput parallel purification of proteins and antibodies. This instrument allows multiple load, wash and elution buffers to be used in parallel along independent lines for up to 24 individual samples. To demonstrate its utility, its use in the purification of five recombinant PB2 C-terminal domains from various subtypes of the influenza A virus is described. Three of these constructs crystallized and one diffracted X-rays to sufficient resolution for structure determination and deposition in the Protein Data Bank. Methods for screening lysis buffers for a cytochrome P450 from a pathogenic fungus prior to upscaling expression and purification are also described. The Protein Maker has become a valuable asset within the Seattle Structural Genomics Center for Infectious Disease (SSGCID) and hence is a potentially valuable tool for a variety of high-throughput protein-purification applications.

摘要

蛋白质制备仪是Emerald生物系统公司开发的一种自动化纯化系统,用于蛋白质和抗体的高通量平行纯化。该仪器允许沿着独立的管路并行使用多种上样、洗涤和洗脱缓冲液,最多可处理24个独立样品。为了展示其效用,本文描述了它在从甲型流感病毒的各种亚型中纯化五个重组PB2 C末端结构域的应用。其中三个构建体结晶,一个衍射出足够分辨率的X射线用于结构测定并提交到蛋白质数据库。本文还描述了在扩大表达和纯化规模之前,筛选来自致病真菌的细胞色素P450裂解缓冲液的方法。蛋白质制备仪已成为西雅图传染病结构基因组学中心(SSGCID)的一项宝贵资产,因此对于各种高通量蛋白质纯化应用而言是一种潜在的有价值工具。

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