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所在平台: Udemy |
课程主页: https://www.udemy.com/course/production-and-purification-of-recombinant-proteins/
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课程名称：重组蛋白的生产与纯化 课程概述： 本课程专注于重组蛋白的生产与纯化，特别是利用大肠杆菌（Escherichia coli）作为表达平台的最佳实践。大肠杆菌因其长期作为细胞工厂的历史，成为重组蛋白生产的首选物种。课程将介绍多种表达质粒、改进的菌株以及多样的培养技术，以实现外源蛋白的高效生产。 在大肠杆菌中，重组蛋白通常以不溶性聚集物（包涵体）的形式产生。这些包涵体蛋白位于低速沉淀分中，可以通过与混合洗涤剂的提取进行部分纯化。重组蛋白的大规模生产中，蛋白质重折叠是一个关键步骤，必须将溶解的/展开的蛋白质重折叠为正确的构象，以便获得具有生物活性的形式。 课程内容涵盖了重组蛋白高产的相关方法学，以及使用柱色谱进行纯化的技术。具体包括发酵的类型、细菌发酵，如何从总细菌内容中提取表达的蛋白质作为不溶性聚集物、蛋白质重折叠的策略、色谱分类的示意图、FPLC和HPLC技术、提高样品纯度的分离能力、根据蛋白质结构的色谱方法类型，以及如何使用SDS-PAGE系统分析分离的蛋白质。 本课程旨在为学员提供重组蛋白生产与纯化的全面方法和实践技巧。
One of the preferred species for the creation of recombinant proteins is Escherichia coli. It is now the most widely used expression platform and has a long history of use as a cell factory. Because of this, a wide range of expression plasmids, a large number of modified strains, and a variety of cultivation techniques are available for the high-level production of heterologous proteins. Recombinant proteins are produced in E. coli as insoluble aggregates called inclusion bodies (IBs). Inclusion body proteins, which are located in the low-speed pellet fraction, can be partially purified by extracting with a mixture of detergentProtein refolding is a key step for large scale production of recombinant proteins. Solubilized/unfolded protein needs to be refolded into the correct conformation to obtain a biologically active form.This course describes the methodology of high production of recombinant proteins and the techniques of purification using column chromatography. The course explains types of fermentation, bacterial fermentation. how can we extract expressed proteins as insoluble aggregates from total bacterial content. Strategy of protein refoldings. Schematic diagram of chromatography classifications. FPLC and HPLC techniques. Separation power for increasing the purity of sample. Types of chromatographic methods according to protein structure. How to analyze separated proteins using SDS-PAGE system.
