High‐Throughput Single‐Cell‐Resolved Spatial Proteomics Enabled by an Ordered Colloidal Crystal Column
, 10-fold higher than that of commonly used sub-2-µm particle packed columns, enabling robust and rapid peptide separation. With such a column, high-throughput nanoLC-MS analysis was achieved, as demonstrated by the identification of 5942 and 4388 proteins from 250 pg HeLa digests using 5- and 2-min gradients, respectively. More notably, the column exhibited exceptional performance in single-cell spatial proteomics, enabling the identification of up to 2304 proteins from a single hepatocyte slice within only a 5-min gradient. Even under an ultrarapid 2-min gradient, up to 1292 proteins were identified from single-cell slices, which is 16 times faster than conventional methods. All these results demonstrated great promise of the colloidal crystal column for high-throughput spatial proteomics with single-cell resolution.
Authors
- Baofeng Zhao (ORCID: https://orcid.org/0000-0002-4920-2661)
- Yu Zhang (ORCID: https://orcid.org/0000-0002-2358-0904)
- Liang Yu (ORCID: https://orcid.org/0009-0007-3922-3454)
- Jianhong Wu (ORCID: https://orcid.org/0000-0003-0052-5336)
- Kun Guo
- Haofei Sun
- Lihua Zhang (ORCID: https://orcid.org/0000-0003-2543-1547)
- Liming Wang (ORCID: https://orcid.org/0000-0003-0914-6113)
- Zhen Liang
- Chao Wang
- Shan Li
Institutions
- Dalian Institute of Chemical Physics (CN)
- Dalian Medical University (CN)
- Second Affiliated Hospital of Dalian Medical University (CN)
- University of Chinese Academy of Sciences (CN)
- Thermo Fisher Scientific (China) (CN)
Publication Details
- Journal
- Angewandte Chemie International Edition
- Published
- 2026-04-27
- DOI
- https://doi.org/10.1002/anie.8045649
- Primary Topic
- Advanced Proteomics Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Natural Science Foundation of China
- Liaoning Revitalization Talents Program
- National Key Research and Development Program of China