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Integrated Proteomic Sample Preparation with Combination of On-line High-abundance Protein Depletion, Denaturation, Reduction, Desalting and Digestion to Achieve High Throughput Plasma Proteome Quantification by Yilan Li; Huiming Yuan; Zhongpeng Dai; Weijie Zhang; Xiaodan Zhang; Baofeng Zhao; Zhen Liang; Lihua Zhang; Yukui Zhang is a Chemistry article available to read on EtoBox.

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In this study, we developed an integrated plasma proteome sample preparation system, by which high-abundance proteins from human plasma were first depleted by immunoaffinity column, followed by on-line middle and low-abundance proteins denaturation, reduction, desalting and tryptic digestion. To evaluate the performance of such a system, 20 μL plasma was processed automatically, followed by 1-h gradient liquid chromatography-mass spectrometry analysis (LC-MS). Compared to conventional in-solution protocols, not only the sample preparation time could be shortened from 20 h to 20 min, but also the number of identified proteins were greatly increased by 1.4–2.0 times. Such an integrated system allows us to process 36 human plasma samples per day, with more than 300 proteins and 52 FDA approved disease markers per sample being identified. With combination of such an integrated sample preparation system with label-free single-shot LC-MS/MS, the human plasma proteins could be quantified across more than 6 orders of magnitude of abundance range with high reproducibility (Pearson R = 0.99, n = 9). In addition, the relative quantification of human plasma samples from diabetic retinopathy pa

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Author

Yilan Li; Huiming Yuan; Zhongpeng Dai; Weijie Zhang; Xiaodan Zhang; Baofeng Zhao; Zhen Liang; Lihua Zhang; Yukui Zhang

Publisher

Elsevier BV

Published

2021

Language

EN

Field

Chemistry (Physical Sciences)