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使用Pichia pastoris中的N终端透皮体来表达和净化复合人体III型原蛋白

Yu Wang^1,2, Ya Feng^1,2, Wansen Tan^1,2

¹School of Life Science and Medical Engineering, Anhui University, Hefei, China.

Current protocols

|December 11, 2025

概括

研究人员开发了一种可扩展的Pichia pastoris发酵工艺,用于重组类型III,实现了高产量,并证明了其对潜在工业应用的生物活性.

科学领域:

生物技术是生物技术.
蛋白质工程是指蛋白质工程.
细胞培养技术细胞培养技术

背景情况:

重组原蛋白的大规模生产对于各种应用至关重要.
优化表达系统和净化方法对于高效的制造至关重要.

研究的目的:

建立一个标准化的技术框架,用于大规模的重组原蛋白生产.
为了优化通过皮肤类型III原融合蛋白 (通过皮肤-hCOL3A) 的发酵和净化过程.

主要方法:

使用Pichia pastoris GS115作为宿主表达系统.
为融合基因设计了一种具有编码子优化的重组载体.
在一个15L生物反应器中进行了高细胞密度的食批发发酵.
优化下游净化使用膜度和阴离子交换色谱.

主要成果:

实现了最终的重组蛋白产量为1.9g/L.
通过优化净化,提升目标蛋白恢复到68%.
经证实通过凝过的透皮-hCOL3A存在于通过凝过的三分剂中.
通过HaCaT细胞增殖试验证明了生物功能.

结论:

成功建立了一个强大的和可扩展的过程,用于再生原蛋白的生产.
为试点规模的制造和未来的工业规模的翻译提供了技术基础.
验证了所产生的复合原III型的生物活性.

关键词:

15L发酵器中的发酵器为15L.这就是Pichia pastoris.净化净化净化净化通过皮肤的化物hCOL3A.

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