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Improved profiling of low molecular weight serum proteome for gastric carcinoma by data-independent acquisition

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Abstract

Low molecular weight proteins (LMWPs) in the bloodstream participate in various biological processes and are closely associated with disease status, whereas identification of serous LMWPs remains a great technical challenge due to the wide dynamic range of protein components. In this study, we constructed an integrated LMWP library by combining the LMWPs obtained by three enrichment methods (50% ACN, 20% ACN + 20 mM ABC, and 30 kDa) and their fractions identified by the data-dependent acquisition method. With this newly constructed library, we comprehensively profiled LMWPs in serum using data-independent acquisition and reliably achieved quantitative results for 75% serous LMWPs. When applying this strategy to quantify LMWPs in human serum samples, we could identify 405 proteins on average per sample, of which 136 proteins were with a MW less than 30 kDa and 293 proteins were with a MW less than 65 kDa. Of note, pre- and post-operative gastric carcinoma (GC) patients showed differentially expressed serous LWMPs, which was also different from the pattern of LWMP expression in healthy controls. In conclusion, our results showed that LMWPs could efficiently distinguish GC patients from healthy controls as well as between pre- and post-operative statuses, and more importantly, our newly developed LMWP profiling platform could be used to discover candidate LMWP biomarkers for disease diagnosis and status monitoring.

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Data availability

Data were deposited to the ProteomeXchange Consortium via PRIDE with the identifier PXD034184.

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Funding

This work received financial support from the National Natural Science Foundation of Guangdong Province (Grant No. 2020A1515010576, No. 2018A0303100023), the National Natural Science Foundation of China (Grant No. 22004086), and the Shenzhen Basic Discipline Layout Project (Grant No. JCYJ20210324115601005).

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Authors and Affiliations

  1. The Central Laboratory, Shenzhen Second People’s Hospital/the First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China

    Weifeng Li, Mengna Li, Xiaoli Zhang, Siqin Yue, Yun Xu, Wenjing Jian & Wenlan Liu

  2. Department of Gastrointestinal Surgery, Shenzhen Second People’s Hospital/the First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China

    Yin Qin

  3. Sustech Core Research Facilities, Southern University of Science and Technology, Shenzhen, 518055, China

    Lin Lin

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  1. Weifeng Li
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Correspondence to Yin Qin, Lin Lin or Wenlan Liu.

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Li, W., Li, M., Zhang, X. et al. Improved profiling of low molecular weight serum proteome for gastric carcinoma by data-independent acquisition. Anal Bioanal Chem 414, 6403–6417 (2022). https://doi.org/10.1007/s00216-022-04196-z

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