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Citation: LIU Jianhui, ZHANG Xiaodan, ZHOU Yuan, SHAN Yichu, FANG Fei, ZHANG Lihua, ZHANG Yukui. A cross-sequence labeling equipment for high-throughput proteome based on pseudo-isobaric dimethyl labeling[J]. Chinese Journal of Chromatography, ;2018, 36(3): 230-236. doi: 10.3724/SP.J.1123.2017.11040 shu

A cross-sequence labeling equipment for high-throughput proteome based on pseudo-isobaric dimethyl labeling

1.
Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;
2.
University of Chinese Academy of Sciences, Beijing 100049, China

Received Date: 27 November 2017

Fund Project: National Natural Science Foundation of China (No. 21405154).

Facing the needs of multidimensional and multi-sample proteome quantification in the field of biology and precision medicine, high-throughput quantitative labeling and analysis have become the trend in recent years. A novel cross-sequence labeling equipment for high-throughput proteome based on pseudo-isobaric dimethyl labeling (pIDL-StageTip) was established. The equipment needs only simple devices and centrifugal force. The throughput of quantitative labeling is effectively increased, and the controllability of the two-step labeling reaction time at the terminals of the peptides and the ease of operation are ensured. By optimizing the labeling conditions of NaBD₃CN and NaBH₃CN under acidic conditions respectively, labeling efficiency of the standard proteolysis achieved 100% and labeling selectivity was more than 95%. Even in the complex human proteome, the labeling efficiency was greater than 99% and the labeling selectivity was 100%. The quantitative method based on the equipment showed high quantitative accuracy and precision. The equipment provides a reliable solution for quantitative proteome labeling with high operability, accuracy, and throughput.
- proteome labeling equipment,
- pseudo-isobaric dimethyl labeling (pIDL),
- cross-sequence labeling using site-selectivity,
- high throughput
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