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An angled-shape tip-based strategy for highly sensitive proteomic profiling of a low number of cells
Profiling proteins plays an essential role in understanding the functions and dynamic networks in biological systems. Mass spectrometry-based proteomic analysis commonly requires multistep sample processing, which results in severe sample loss. Although the recently developed microproteomic strategi...
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Published in: | Analytical methods 2023-03, Vol.15 (9), p.1215-1222 |
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creator | Zhao, Hongxian Chen, Yongle Li, Hang Zhang, Yangjun Zhang, Wanjun Qin, Weijie |
description | Profiling proteins plays an essential role in understanding the functions and dynamic networks in biological systems. Mass spectrometry-based proteomic analysis commonly requires multistep sample processing, which results in severe sample loss. Although the recently developed microproteomic strategies have substantially reduced sample loss
droplet microfluidic technology, specialized equipment and well-trained personnel are needed, which may limit their wide adoption. Here, we report an angled-shape tip-based strategy for rapid sample preparation and sensitive proteomic profiling of small cell populations ( |
doi_str_mv | 10.1039/d2ay01884e |
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droplet microfluidic technology, specialized equipment and well-trained personnel are needed, which may limit their wide adoption. Here, we report an angled-shape tip-based strategy for rapid sample preparation and sensitive proteomic profiling of small cell populations (<1000 cells). The angled-shape tip provided a 'reactor' for the entire proteomic sample processing workflow, from cell capture and lysis to protein digestion, eliminating the sample transfer-induced protein loss. The angled-shape tip was surface-treated for anti-protein adsorption which further reduced the sample loss. Using this strategy, 1241 ± 38-4110 ± 37 protein groups and 4010 ± 700-34 879 ± 575 peptides were identified from 10-1000 HeLa cells with high quantification reproducibility in only 4.5 h sample processing time, which was superior to the reported methods and commercial kits, especially for <100 cells. This approach was easily accessible, straightforward to operate, and compatible with flow cytometry-based cell sorting. It showed great potential for in-depth proteomic profiling of rare cells (<1000 cells) in both basic biological research and clinical application.</description><identifier>ISSN: 1759-9660</identifier><identifier>EISSN: 1759-9679</identifier><identifier>DOI: 10.1039/d2ay01884e</identifier><identifier>PMID: 36804579</identifier><language>eng</language><publisher>England: Royal Society of Chemistry</publisher><subject>Biological research ; Flow cytometry ; HeLa Cells ; Humans ; Lysis ; Mass spectrometry ; Mass spectroscopy ; Microfluidics ; Peptides ; Protein adsorption ; Proteins ; Proteins - analysis ; Proteomics ; Proteomics - methods ; Reproducibility of Results ; Sample preparation ; Workflow</subject><ispartof>Analytical methods, 2023-03, Vol.15 (9), p.1215-1222</ispartof><rights>Copyright Royal Society of Chemistry 2023</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c274t-2debc347e8130529b07f2a4ece79c1acad96650ff3290847b74058b097fb08063</cites><orcidid>0000-0002-7633-9786 ; 0000-0002-7631-6808</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,786,790,27957,27958</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36804579$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Hongxian</creatorcontrib><creatorcontrib>Chen, Yongle</creatorcontrib><creatorcontrib>Li, Hang</creatorcontrib><creatorcontrib>Zhang, Yangjun</creatorcontrib><creatorcontrib>Zhang, Wanjun</creatorcontrib><creatorcontrib>Qin, Weijie</creatorcontrib><title>An angled-shape tip-based strategy for highly sensitive proteomic profiling of a low number of cells</title><title>Analytical methods</title><addtitle>Anal Methods</addtitle><description>Profiling proteins plays an essential role in understanding the functions and dynamic networks in biological systems. Mass spectrometry-based proteomic analysis commonly requires multistep sample processing, which results in severe sample loss. Although the recently developed microproteomic strategies have substantially reduced sample loss
droplet microfluidic technology, specialized equipment and well-trained personnel are needed, which may limit their wide adoption. Here, we report an angled-shape tip-based strategy for rapid sample preparation and sensitive proteomic profiling of small cell populations (<1000 cells). The angled-shape tip provided a 'reactor' for the entire proteomic sample processing workflow, from cell capture and lysis to protein digestion, eliminating the sample transfer-induced protein loss. The angled-shape tip was surface-treated for anti-protein adsorption which further reduced the sample loss. Using this strategy, 1241 ± 38-4110 ± 37 protein groups and 4010 ± 700-34 879 ± 575 peptides were identified from 10-1000 HeLa cells with high quantification reproducibility in only 4.5 h sample processing time, which was superior to the reported methods and commercial kits, especially for <100 cells. This approach was easily accessible, straightforward to operate, and compatible with flow cytometry-based cell sorting. It showed great potential for in-depth proteomic profiling of rare cells (<1000 cells) in both basic biological research and clinical application.</description><subject>Biological research</subject><subject>Flow cytometry</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Lysis</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Microfluidics</subject><subject>Peptides</subject><subject>Protein adsorption</subject><subject>Proteins</subject><subject>Proteins - analysis</subject><subject>Proteomics</subject><subject>Proteomics - methods</subject><subject>Reproducibility of Results</subject><subject>Sample preparation</subject><subject>Workflow</subject><issn>1759-9660</issn><issn>1759-9679</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><recordid>eNpdkEtPwzAQhC0EoqVw4QcgS1wQUmCdl-1jVcpDqsQFDpwiJ1mnrvIodgLKvyehpQf2srPSp9HsEHLJ4I5BIO9zX_XAhAjxiEwZj6QnYy6PDzqGCTlzbgMQyyBmp2QSxALCiMspyec1VXVRYu65tdoibc3WS5XDnLrWqhaLnurG0rUp1mVPHdbOtOYL6dY2LTaVyUalTWnqgjaaKlo237TuqhTteGdYlu6cnGhVOrzY7xl5f1y-LZ691evTy2K-8jKfh63n55hmQchRsAAiX6bAta9CzJDLjKlM5cMvEWgd-BJEyFMeQiRSkFynICAOZuRm5ztE-uzQtUll3JhA1dh0LvE5F5JHIuIDev0P3TSdrYd0AyXYMALkQN3uqMw2zlnUydaaStk-YZCM3ScP_vzjt_vlAF_tLbu0wvyA_pUd_ABgA36Y</recordid><startdate>20230302</startdate><enddate>20230302</enddate><creator>Zhao, Hongxian</creator><creator>Chen, Yongle</creator><creator>Li, Hang</creator><creator>Zhang, Yangjun</creator><creator>Zhang, Wanjun</creator><creator>Qin, Weijie</creator><general>Royal Society of Chemistry</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SE</scope><scope>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>FR3</scope><scope>H8G</scope><scope>JG9</scope><scope>L7M</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-7633-9786</orcidid><orcidid>https://orcid.org/0000-0002-7631-6808</orcidid></search><sort><creationdate>20230302</creationdate><title>An angled-shape tip-based strategy for highly sensitive proteomic profiling of a low number of cells</title><author>Zhao, Hongxian ; Chen, Yongle ; Li, Hang ; Zhang, Yangjun ; Zhang, Wanjun ; Qin, Weijie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c274t-2debc347e8130529b07f2a4ece79c1acad96650ff3290847b74058b097fb08063</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Biological research</topic><topic>Flow cytometry</topic><topic>HeLa Cells</topic><topic>Humans</topic><topic>Lysis</topic><topic>Mass spectrometry</topic><topic>Mass spectroscopy</topic><topic>Microfluidics</topic><topic>Peptides</topic><topic>Protein adsorption</topic><topic>Proteins</topic><topic>Proteins - analysis</topic><topic>Proteomics</topic><topic>Proteomics - methods</topic><topic>Reproducibility of Results</topic><topic>Sample preparation</topic><topic>Workflow</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, Hongxian</creatorcontrib><creatorcontrib>Chen, Yongle</creatorcontrib><creatorcontrib>Li, Hang</creatorcontrib><creatorcontrib>Zhang, Yangjun</creatorcontrib><creatorcontrib>Zhang, Wanjun</creatorcontrib><creatorcontrib>Qin, Weijie</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Copper Technical Reference Library</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Hongxian</au><au>Chen, Yongle</au><au>Li, Hang</au><au>Zhang, Yangjun</au><au>Zhang, Wanjun</au><au>Qin, Weijie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An angled-shape tip-based strategy for highly sensitive proteomic profiling of a low number of cells</atitle><jtitle>Analytical methods</jtitle><addtitle>Anal Methods</addtitle><date>2023-03-02</date><risdate>2023</risdate><volume>15</volume><issue>9</issue><spage>1215</spage><epage>1222</epage><pages>1215-1222</pages><issn>1759-9660</issn><eissn>1759-9679</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>Profiling proteins plays an essential role in understanding the functions and dynamic networks in biological systems. Mass spectrometry-based proteomic analysis commonly requires multistep sample processing, which results in severe sample loss. Although the recently developed microproteomic strategies have substantially reduced sample loss
droplet microfluidic technology, specialized equipment and well-trained personnel are needed, which may limit their wide adoption. Here, we report an angled-shape tip-based strategy for rapid sample preparation and sensitive proteomic profiling of small cell populations (<1000 cells). The angled-shape tip provided a 'reactor' for the entire proteomic sample processing workflow, from cell capture and lysis to protein digestion, eliminating the sample transfer-induced protein loss. The angled-shape tip was surface-treated for anti-protein adsorption which further reduced the sample loss. Using this strategy, 1241 ± 38-4110 ± 37 protein groups and 4010 ± 700-34 879 ± 575 peptides were identified from 10-1000 HeLa cells with high quantification reproducibility in only 4.5 h sample processing time, which was superior to the reported methods and commercial kits, especially for <100 cells. This approach was easily accessible, straightforward to operate, and compatible with flow cytometry-based cell sorting. It showed great potential for in-depth proteomic profiling of rare cells (<1000 cells) in both basic biological research and clinical application.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>36804579</pmid><doi>10.1039/d2ay01884e</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0002-7633-9786</orcidid><orcidid>https://orcid.org/0000-0002-7631-6808</orcidid></addata></record> |
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subjects | Biological research Flow cytometry HeLa Cells Humans Lysis Mass spectrometry Mass spectroscopy Microfluidics Peptides Protein adsorption Proteins Proteins - analysis Proteomics Proteomics - methods Reproducibility of Results Sample preparation Workflow |
title | An angled-shape tip-based strategy for highly sensitive proteomic profiling of a low number of cells |
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