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Selective flocculation and precipitation for the improvement of virus-like particle recovery from yeast homogenate

机译:选择性絮凝和沉淀可改善酵母匀浆中病毒样颗粒的回收率

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摘要

The purification of an intracellular product from a complex mixture of contaminants after cell disruption is a common problem in processes downstream of fermentation systems. This is particularly challenging for the recovery of particulate (80 nm indiameter) multimeric protein products, named virus-like particles (VLPs), from cell debris and other intracellular components. Selective flocculation for debris removal followed by selective precipitation of the target protein can be used as a preclarification step to aid purification. In this paper, selective borax flocculation of cell debris in yeast homogenate, followed by selective poly (ethylene glycol) precipitation of VLPs are defined with a view to demonstrating their potential in aiding the initial clarification stages of the purification sequence. The translation from laboratory scale to pilot scale operation is addressed, demonstrating the challenge of scale-up of solid - liquid separation stages for biological particle processing.
机译:细胞分裂后从复杂的污染物混合物中纯化细胞内产物是发酵系统下游过程中的普遍问题。这对于从细胞碎片和其他细胞内组分中回收称为病毒样颗粒(VLP)的颗粒(直径80 nm)多聚体蛋白质产品而言尤其具有挑战性。选择性絮凝去除杂物,然后选择性沉淀目标蛋白可以用作帮助纯化的预纯化步骤。在本文中,定义了酵母匀浆中细胞碎片的选择性硼砂絮凝,然后选择性地沉淀VLP的聚(乙二醇),以展示其在协助纯化序列初始澄清阶段中的潜力。解决了从实验室规模到中试规模的转换,这说明了用于生物颗粒处理的固液分离阶段规模扩大的挑战。

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