利用丝状球化技术实时监测重组GFP在法菲Komagataella phaffii单细胞中的表达

IF 3.6 3区化学 Q2 CHEMISTRY, ANALYTICAL

Reviews in Analytical Chemistry Pub Date : 2023-01-01 DOI:10.1515/revac-2023-0059

Xiao Zheng, Wenjie Cong, Hualan Zhou, Jianguo Zhang

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引用次数: 0

摘要

摘要法菲Komagataella phaffii是甲醇诱导下重组蛋白生产的重要细胞工厂。追踪重组蛋白在单个菲氏k细胞中的表达是确定其在潜培养系统中表达水平变化的一个热点。本研究建立了一种利用黑曲霉微球固定法菲氏k细胞的平台系统，利用荧光显微镜检测单细胞绿色荧光蛋白(GFP)的表达。通过微球制备、K. phaffii细胞吸收、诱导剂比较、甲醇浓度、介质流速等因素对该体系进行优化，获得了一个成功的追踪单个K. phaffii细胞GFP表达的平台。该系统提供了一种在线分析技术来发现各种菲氏k细胞的异质性能力。

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Real-time monitoring of recombinant GFP expression in single-cell Komagataella phaffii through filamentous pelletization technology

Abstract Komagataella phaffii is an important cell factory for recombinant protein production under methanol induction. Tracking of recombinant protein expression by single K. phaffii cell is a hot topic to identify the variation of expression level in submerge cultivation system. In this study, a platform system harnessing Aspergillus niger pellets for K. phaffii cell immobilization in fabricated glass plate was developed to measure the green fluorescent protein (GFP) expression of single cell using fluorescence microscope. This system was optimized through pellets preparation, K. phaffii cell absorption, inducer comparison, methanol concentration, flow velocity of medium, and obtaining a successful platform to track GFP expression of single K. phaffii cell. This system provided an on-line analytical technology to discover the heterogeneous capability of various K. phaffii cells.

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来源期刊

Reviews in Analytical Chemistry 化学-分析化学

CiteScore

7.50

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Reviews in Analytical Chemistry publishes authoritative reviews by leading experts in the dynamic field of chemical analysis. The subjects can encompass all branches of modern analytical chemistry such as spectroscopy, chromatography, mass spectrometry, electrochemistry and trace analysis and their applications to areas such as environmental control, pharmaceutical industry, automation and other relevant areas. Review articles bring the expert up to date in a concise manner and provide researchers an overview of new techniques and methods.