Effect of pre-filtration on selective isolation of tat protein by affinity membrane separation: Analysis of flux, separation efficiency, and processing time
Effect of pre-filtration on selective isolation of tat protein by affinity membrane separation: Analysis of flux, separation efficiency, and processing time
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DOI：
10.1080/01496390701477121
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发表时间：
2007-01-01
影响因子：
2.8
通讯作者：
Toborek, M.
中科院分区：
文献类型：
--
作者：
Datta, Saurav;Bhattacharyya, D.;Toborek, M.
文献中报道了在微滤膜中使用抗生物素蛋白-生物素相互作用从细菌裂解物中分离和纯化达特蛋白。为了提高该技术的效率，必须改进通量、达特分离效率和处理时间。在目前的研究工作中，引入了预过滤步骤，以在亲和膜分离之前从进料中除去不需要的高分子量蛋白质和其他杂质。观察到达特的通量和分离效率的显著增强。处理时间也大大缩短。例如，使用UF预处理步骤，总达特回收率比使用未处理的进料所观察到的高约4倍（处理时间低25%）。纯化后的达特蛋白经SDS-PAGE、Western Blot和生物素分析鉴定。用模型方程描述了亲和分离中的通量行为。
The isolation and purification of Tat protein from bacterial lysate using avidinbiotin interaction in microfiltration membranes have been reported in the literature. To increase the efficacy of the technique, improvements in flux, Tat separation efficiency, and processing time are essential. In the current research work a pre-filtration step was introduced to remove unwanted high molecular weight proteins and other impurities from feed prior to affinity membrane separation. Significant enhancement in flux and separation efficiency of Tat was observed. Processing time was also reduced significantly. For example, with UF pretreatment step the total Tat recovery was around four times higher (with processing time 25% lower) than that observed with the untreated feed. The quality of purified Tat was analyzed by SDS-PAGE, Western Blot, and biotin analysis. Flux behavior in affinity separation was described by model equations.