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首页> 外文期刊>Biotechnology Progress >Initial Evaluation of Protein Throughput and Yield Characteristics on Nylon 6 Capillary-Channeled Polymer (C-CP) Fiber Stationary Phases by Frontal Analysis
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Initial Evaluation of Protein Throughput and Yield Characteristics on Nylon 6 Capillary-Channeled Polymer (C-CP) Fiber Stationary Phases by Frontal Analysis

机译：额叶分析法初步评估尼龙6毛细通道聚合物（C-CP）纤维固定相的蛋白质通量和产率特性

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摘要

Nylon 6 capillary-channeled polymer (C-CP) fibers are investigated as an alternative support! stationary phase for downstream processing of macromolecules. Ionizable amine and carboxylic acid end groups on the native fiber surface allow for ion exchange chromatography (IEC). The low cost and ability to operate at high linear velocities and low back pressures are practical advantages of C-CP fibers for preparative-scale macromolecule separations. The lack of fiber porosity ensures facile adsorption/ desorption that is conducive to high throughput and recoveries/yields. Described here is a preliminary investigation of the processing characteristics of lysozyme on nylon 6 fibers with an eye toward downstream processing applications. Fibers were packed into microbore (0.8 mm i.d.) and analytical-size (2.1 mm i.d.) columns for the evaluation of the role of linear velocity on pressure drop, frontal throughput, and yield. Protein isolation by frontal development involved three steps: loading of the column to breakthrough, an aqueous wash, and a salt wash to recover the protein. Frontal throughput was evaluated with different salt concentrations (0-1000 mM NaCl) and different linear velocities (6—24 mm s~(-1) ). The observed throughput values are in the range of 0.12-0.20 mg min~(-1) when 0.25 mg mL~(-1) lysozyme (in 20 mM Tris-HCl) is loaded onto 78 mg of C-CP fiber in 0.52 mL volume analytical columns. Increased throughput and yield were found when protein was loaded and eluted at high linear velocity. Results of this study lend credence to the further development of C-CP fibers for biomacromolecule processing on larger scales.
机译：研究了尼龙6毛细管聚合物（C-CP）纤维作为替代支撑物！固定相，用于大分子的下游加工。天然纤维表面的可电离胺和羧酸端基可用于离子交换色谱（IEC）。 C-CP纤维的低成本和在高线速度下运行的能力以及低的背压是C-CP纤维用于制备规模大分子分离的实际优势。纤维孔隙率的缺乏确保了容易的吸附/解吸，有利于高通量和回收率/收率。这里介绍的是尼龙6纤维上溶菌酶的加工特性的初步研究，着眼于下游加工应用。将纤维填充到微孔（内径0.8毫米）和分析尺寸（内径2.1毫米）的色谱柱中，以评估线速度对压降，正面通量和产量的作用。通过额叶发育进行蛋白质分离包括三个步骤：将色谱柱上样至突破，水洗和盐洗以回收蛋白质。用不同的盐浓度（0-1000 mM NaCl）和不同的线速度（6-24 mm s〜（-1））评估额叶吞吐量。当将0.25 mg mL〜（-1）溶菌酶（在20 mM Tris-HCl中）加载到0.52 mL中的78 mg C-CP纤维上时，观察到的通量值在0.12-0.20 mg min〜（-1）的范围内体积分析柱。当蛋白质以高线速度上样和洗脱时，发现产量和产量都有提高。这项研究的结果为大规模生产生物大分子的C-CP纤维的进一步开发提供了保证。

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