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芯片过器:微流体基础的微生物群体综合样本准备方法

Ranjith Kumar Ravi Kumar¹, Massamba Mbacke Ndiaye¹, Iman Haddad¹

¹Spectrométrie de Masse Biologique et Protéomique, LPC, UMR ESPCI CNRS 8249, 10 rue Vauquelin, F-75005 Paris, France.

Journal of proteome research

|February 14, 2024

概括

一种新的微流体ChipFilter设备使细胞直接溶解和蛋白质消化能够用于metaproteomics. 这种方法增强了微生物蛋白质的识别,即使是从低密度样本,改善微生物组的功能分析.

科学领域:

微生物学微生物学
蛋白质组学是指蛋白质组学.
分析化学分析化学

背景情况:

超蛋白质组学提供了对微生物组的功能性见解,但在从复杂样本中提取蛋白质方面面临着挑战.
目前的方法与低细胞密度和异质微生物群落作斗争.

研究的目的:

评估ChipFilter微流体装置用于直接微生物细胞溶解和蛋白质消化在metaproteomic工作流程中.
评估ChipFilter与LC-MS/MS结合用于识别微生物蛋白质的效率.

主要方法:

使用ChipFilter微流体装置进行直接细胞溶解和现场蛋白质消化.
采用液体染色学并联质谱法 (LC-MS/MS) 用于的识别和量化.
用纯种植物 (大肠杆菌,B. subtilis,S. cerevisiae) 和复杂的17种肠道微生物组混合物测试了系统.

主要成果:

芯片波器系统成功地从溶解细胞中直接识别出微生物蛋白质.
与标准协议相比,发现了更多的蛋白质和,特别是在细胞密度低的情况下.
在复杂的肠道微生物样本中,ChipFilter制剂识别和量化了17种物种中的16种.

结论:

芯片过器微流体装置是微生物组的元蛋白质组分析的有效工具.
这种方法克服了蛋白质提取的技术挑战,特别是对于体积小或细胞密度低的样本.
该方法有助于改善微生物群落的功能性特征.

关键词:

这里是metaproteomic的一个例子.蛋白质基质的保护性样品的准备样品的准备

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相关概念视频

iChip

iChip

The cultivation of environmental microorganisms has long been hindered by the inability to replicate complex native conditions in vitro. The isolation chip (iChip) addresses this limitation by facilitating the growth of previously uncultivable microorganisms through in situ incubation. Designed for high-throughput microbial cultivation, the iChip comprises hundreds of microchambers, each capable of housing a single microbial cell. These microchambers are loaded with a mixture of molten agar and...