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细胞-蛋白质混合物造成的膜污染:利用多光子显微镜进行原位表征

Membrane fouling by cell-protein mixtures: in situ characterisation using multi-photon microscopy.

作者信息

Hughes David J, Cui Zhanfeng, Field Robert W, Tirlapur Uday K

机构信息

Department of Engineering Science, University of Oxford, Parks Road, Oxford OX1 3PJ, UK.

出版信息

Biotechnol Bioeng. 2007 Apr 15;96(6):1083-91. doi: 10.1002/bit.21113.

Abstract

Fouling of the membrane by cell and protein mixtures can result in severe flux declines, leading to the eventual need to clean or replace the membrane. In this study multi-photon microscopy, a fluorescence-based technique is used to 3-D image in situ the fouling of microfiltration membranes by suspensions containing combinations of washed yeast, bovine serum albumin (BSA) and ovalbumin. Appropriate fluorescent labelling allows the three foulant species to be clearly identified. Images correlate well with filtration data and clearly show the cake of yeast cells capturing protein aggregates. The proteins exhibited very different filtration behaviour. When filtering washed yeast together with ovalbumin and/or a 50:50 mixture by mass of BSA and ovalbumin, the ovalbumin fouling dominates the system. Capture of aggregates by the cake did not reduce fouling of the membrane by the protein and increased the resistance of the cake. For mixtures of BSA and washed yeast, the presence of a cake of yeast cells did reduce fouling of the membrane by the protein, however, the extra resistance due to the cake resulted in a flux lower than that when filtering BSA alone.

摘要

细胞与蛋白质混合物对膜的污染会导致通量严重下降,最终需要对膜进行清洗或更换。在本研究中,多光子显微镜(一种基于荧光的技术)用于对含有洗涤过的酵母、牛血清白蛋白(BSA)和卵清蛋白混合物的悬浮液对微滤膜的污染进行原位三维成像。适当的荧光标记可使三种污染物清晰可辨。图像与过滤数据高度相关,清楚地显示了酵母细胞滤饼捕获蛋白质聚集体的情况。这些蛋白质表现出非常不同的过滤行为。当将洗涤过的酵母与卵清蛋白和/或质量比为50:50的BSA与卵清蛋白混合物一起过滤时,卵清蛋白污染主导了整个体系。滤饼对聚集体的捕获并未降低蛋白质对膜的污染,反而增加了滤饼的阻力。对于BSA与洗涤过的酵母的混合物,酵母细胞滤饼的存在确实降低了蛋白质对膜的污染,然而,滤饼带来的额外阻力导致通量低于单独过滤BSA时的通量。

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