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<p>Rapid and comprehensive analysis of complex proteomes across large sample sets is vital for unlocking the potential of systems biology. We present a high-throughput mass spectrometry (MS) proteomics method that integrates narrow-window data-independent acquisition (nDIA) with short-gradient micro-flow chromatography, enabling profiling of &gt;240 samples per day. This optimized MS approach identifies 6,201 and 7,466 human proteins with 1- and 2-min gradients, respectively. As a practical application, we analyzed 507 samples composed of 13 different tissues from mice treated with the enzyme-drug L-asparaginase (ASNase) or its glutaminase-free Q59L mutant, generating a quantitative profile of 11,472 proteins following drug treatment. The MS results confirmed the impact of ASNase on amino acid metabolism in solid tissues. Further analysis revealed broad suppression of anticoagulants and cholesterol metabolism and uncovered numerous tissue-specific dysregulated pathways. In summary, the optimized high-throughput proteomics method accelerates systems-level analysis of a preclinical model to generate biological insights and clinically actionable hypotheses.</p>

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High-throughput multi-organ proteomics workflow for drug efficacy and toxicity analysis

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年2月9日

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  • Yun Xiong,
  • Lin Tan,
  • Wai-kin Chan,
  • Dandan Zhu,
  • Huimin Zhang,
  • Eric S. Yin,
  • Sri Ramya Donepudi,
  • Jibin Ding,
  • Bo Wei,
  • Bao Tran,
  • Sara Martinez,
  • Iqbal Mahmud,
  • Faiza Hanif Waghu,
  • Hamish I. Stewart,
  • Daniel J. Hermanson,
  • Rehan Akbani,
  • John N. Weinstein,
  • Junjie Chen,
  • Philip L. Lorenzi

摘要

Rapid and comprehensive analysis of complex proteomes across large sample sets is vital for unlocking the potential of systems biology. We present a high-throughput mass spectrometry (MS) proteomics method that integrates narrow-window data-independent acquisition (nDIA) with short-gradient micro-flow chromatography, enabling profiling of >240 samples per day. This optimized MS approach identifies 6,201 and 7,466 human proteins with 1- and 2-min gradients, respectively. As a practical application, we analyzed 507 samples composed of 13 different tissues from mice treated with the enzyme-drug L-asparaginase (ASNase) or its glutaminase-free Q59L mutant, generating a quantitative profile of 11,472 proteins following drug treatment. The MS results confirmed the impact of ASNase on amino acid metabolism in solid tissues. Further analysis revealed broad suppression of anticoagulants and cholesterol metabolism and uncovered numerous tissue-specific dysregulated pathways. In summary, the optimized high-throughput proteomics method accelerates systems-level analysis of a preclinical model to generate biological insights and clinically actionable hypotheses.