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通过多参数流式细胞术监测和定量大肠杆菌中包涵体的形成

Monitoring and quantification of inclusion body formation in Escherichia coli by multi-parameter flow cytometry.

作者信息

Wållberg Fredrik, Sundström Helene, Ledung Erika, Hewitt Christopher J, Enfors Sven-Olof

机构信息

Department of Biotechnology, Royal Institute of Technology, Roslagstullsbacken 21, S-10691 Stockholm, Sweden.

出版信息

Biotechnol Lett. 2005 Jul;27(13):919-26. doi: 10.1007/s10529-005-7184-6.

Abstract

Multi-parameter flow cytometry was used to monitor the formation of promegapoietin (PMP) inclusion bodies during a high cell density Escherichia coli fed-batch fermentation process. Inclusion bodies were labelled with a primary antibody and then with a secondary fluorescent antibody. Using this method it was possible to detect PMP inclusion body formation with a high specificity and it was possible to monitor the increased accumulation of the protein with process time (6-48 mg PMP/g CDW) whilst highlighting population heterogeneity.

摘要

在高细胞密度大肠杆菌补料分批发酵过程中,使用多参数流式细胞术监测前巨核细胞生成素(PMP)包涵体的形成。包涵体先用一抗标记,然后用二抗荧光抗体标记。使用这种方法,可以高特异性地检测PMP包涵体的形成,并且可以监测随着发酵时间(6 - 48 mg PMP/g CDW)蛋白质积累的增加,同时突出群体异质性。

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