Optimized Multi-Attribute Method Workflow Addressing Missed Cleavages and Chromatographic Tailing/Carry-Over of Hydrophobic Peptides
Peptide mapping by liquid chromatography mass spectrometry (LC-MS) and the related multi-attribute method (MAM) are well-established analytical tools for verification of the primary structure and mapping/quantitation of co- and post-translational modifications (PTMs) or product quality attributes in...
Gespeichert in:
| Veröffentlicht in: | Analytical chemistry (Washington) 2022-12, Vol.94 (49), p.17195-17204 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 17204 |
|---|---|
| container_issue | 49 |
| container_start_page | 17195 |
| container_title | Analytical chemistry (Washington) |
| container_volume | 94 |
| creator | Kristensen, Dan B. Ørgaard, Martin Sloth, Trine M. Christoffersen, Nanna S. Leth-Espensen, Katrine Z. Jensen, Pernille F. |
| description | Peptide mapping by liquid chromatography mass spectrometry (LC-MS) and the related multi-attribute method (MAM) are well-established analytical tools for verification of the primary structure and mapping/quantitation of co- and post-translational modifications (PTMs) or product quality attributes in biopharmaceutical development. Proteolytic digestion is a key step in peptide mapping workflows, which traditionally is labor-intensive, involving multiple manual steps. Recently, simple high-temperature workflows with automatic digestion were introduced, which facilitate robustness and reproducibility across laboratories. Here, a modified workflow with an automatic digestion step is presented, which includes a two-step digestion at high and low temperatures, as opposed to the original one-step digestion at a high temperature. The new automatic digestion workflow significantly reduces the number of missed cleavages, obtaining a more complete digestion profile. In addition, we describe how chromatographic peak tailing and carry-over is dramatically reduced for hydrophobic peptides by switching from the traditional C18 reversed-phase (RP) column chemistry used for peptide mapping to a less retentive C4 column chemistry. No negative impact is observed on MS/MS-derived sequence coverage when switching to a C4 column chemistry. Overall, the new peptide mapping workflow significantly reduces the number of missed cleavages, yielding more robust and simple data interpretation, while providing dramatically reduced tailing and carry-over of hydrophobic peptides. |
| doi_str_mv | 10.1021/acs.analchem.2c03820 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_9753059</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2757124129</sourcerecordid><originalsourceid>FETCH-LOGICAL-a477t-c0591e2cd16533b394e8fa647dc6fdbe5fbf34691e08852a03f88f06d8e0b4b83</originalsourceid><addsrcrecordid>eNp9kU-P0zAQxS0EYsvCN0AoEhcu6Y7t_L0gVdWyi7RVOSziaDn2pPGSxMFOisp5PziO2q2AA6eRNb_3nkePkLcUlhQYvZLKL2UvW9Vgt2QKeMHgGVnQlEGcFQV7ThYAwGOWA1yQV94_AFAKNHtJLnjGk6wEuiCP22E0nfmFOtpM7Wji1Tg6U00jRhscG6ujb9Z9r1v7M1pp7dB70--ijfE-KNYtyr3coY9kH16Ns50c7c7JoTEqupemDfDVWjp3iLd7dJGto9uDdnZobBWILxjCNfrX5EUtW49vTvOSfP10fb--je-2N5_Xq7tYJnk-xgrSkiJTmmYp5xUvEyxqmSW5VlmtK0zrqp7voghFkTIJvC6KGjJdIFRJVfBL8vHoO0xVh1phPzrZisGZTrqDsNKIvze9acTO7kWZpzyEB4MPJwNnf0zoR9EZr7BtZY928oLlIZ8yVvKAvv8HfbCTC33NVJpTllA2GyZHSjnrvcP6_BkKYq5ZhJrFU83iVHOQvfvzkLPoqdcAwBGY5efg_3r-Bk2VumQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2757124129</pqid></control><display><type>article</type><title>Optimized Multi-Attribute Method Workflow Addressing Missed Cleavages and Chromatographic Tailing/Carry-Over of Hydrophobic Peptides</title><source>MEDLINE</source><source>American Chemical Society Journals</source><creator>Kristensen, Dan B. ; Ørgaard, Martin ; Sloth, Trine M. ; Christoffersen, Nanna S. ; Leth-Espensen, Katrine Z. ; Jensen, Pernille F.</creator><creatorcontrib>Kristensen, Dan B. ; Ørgaard, Martin ; Sloth, Trine M. ; Christoffersen, Nanna S. ; Leth-Espensen, Katrine Z. ; Jensen, Pernille F.</creatorcontrib><description>Peptide mapping by liquid chromatography mass spectrometry (LC-MS) and the related multi-attribute method (MAM) are well-established analytical tools for verification of the primary structure and mapping/quantitation of co- and post-translational modifications (PTMs) or product quality attributes in biopharmaceutical development. Proteolytic digestion is a key step in peptide mapping workflows, which traditionally is labor-intensive, involving multiple manual steps. Recently, simple high-temperature workflows with automatic digestion were introduced, which facilitate robustness and reproducibility across laboratories. Here, a modified workflow with an automatic digestion step is presented, which includes a two-step digestion at high and low temperatures, as opposed to the original one-step digestion at a high temperature. The new automatic digestion workflow significantly reduces the number of missed cleavages, obtaining a more complete digestion profile. In addition, we describe how chromatographic peak tailing and carry-over is dramatically reduced for hydrophobic peptides by switching from the traditional C18 reversed-phase (RP) column chemistry used for peptide mapping to a less retentive C4 column chemistry. No negative impact is observed on MS/MS-derived sequence coverage when switching to a C4 column chemistry. Overall, the new peptide mapping workflow significantly reduces the number of missed cleavages, yielding more robust and simple data interpretation, while providing dramatically reduced tailing and carry-over of hydrophobic peptides.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.2c03820</identifier><identifier>PMID: 36346901</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Amino acid sequence ; Analytical chemistry ; Biopharmaceuticals ; Chemistry ; Chromatography ; Chromatography, Liquid - methods ; Data interpretation ; Digestion ; High temperature ; Hydrophobicity ; Liquid chromatography ; Low temperature ; Mass spectrometry ; Mass spectroscopy ; Peptide mapping ; Peptide Mapping - methods ; Peptides ; Peptides - chemistry ; Post-translation ; Proteolysis ; Quality management ; Quantitation ; Reproducibility of Results ; Switching ; Tailings ; Tandem Mass Spectrometry - methods ; Workflow</subject><ispartof>Analytical chemistry (Washington), 2022-12, Vol.94 (49), p.17195-17204</ispartof><rights>2022 The Authors. Published by American Chemical Society</rights><rights>Copyright American Chemical Society Dec 13, 2022</rights><rights>2022 The Authors. Published by American Chemical Society 2022 The Authors</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a477t-c0591e2cd16533b394e8fa647dc6fdbe5fbf34691e08852a03f88f06d8e0b4b83</citedby><cites>FETCH-LOGICAL-a477t-c0591e2cd16533b394e8fa647dc6fdbe5fbf34691e08852a03f88f06d8e0b4b83</cites><orcidid>0000-0002-5491-1665</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.2c03820$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.2c03820$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,315,781,785,886,2766,27080,27928,27929,56742,56792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36346901$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kristensen, Dan B.</creatorcontrib><creatorcontrib>Ørgaard, Martin</creatorcontrib><creatorcontrib>Sloth, Trine M.</creatorcontrib><creatorcontrib>Christoffersen, Nanna S.</creatorcontrib><creatorcontrib>Leth-Espensen, Katrine Z.</creatorcontrib><creatorcontrib>Jensen, Pernille F.</creatorcontrib><title>Optimized Multi-Attribute Method Workflow Addressing Missed Cleavages and Chromatographic Tailing/Carry-Over of Hydrophobic Peptides</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Peptide mapping by liquid chromatography mass spectrometry (LC-MS) and the related multi-attribute method (MAM) are well-established analytical tools for verification of the primary structure and mapping/quantitation of co- and post-translational modifications (PTMs) or product quality attributes in biopharmaceutical development. Proteolytic digestion is a key step in peptide mapping workflows, which traditionally is labor-intensive, involving multiple manual steps. Recently, simple high-temperature workflows with automatic digestion were introduced, which facilitate robustness and reproducibility across laboratories. Here, a modified workflow with an automatic digestion step is presented, which includes a two-step digestion at high and low temperatures, as opposed to the original one-step digestion at a high temperature. The new automatic digestion workflow significantly reduces the number of missed cleavages, obtaining a more complete digestion profile. In addition, we describe how chromatographic peak tailing and carry-over is dramatically reduced for hydrophobic peptides by switching from the traditional C18 reversed-phase (RP) column chemistry used for peptide mapping to a less retentive C4 column chemistry. No negative impact is observed on MS/MS-derived sequence coverage when switching to a C4 column chemistry. Overall, the new peptide mapping workflow significantly reduces the number of missed cleavages, yielding more robust and simple data interpretation, while providing dramatically reduced tailing and carry-over of hydrophobic peptides.</description><subject>Amino acid sequence</subject><subject>Analytical chemistry</subject><subject>Biopharmaceuticals</subject><subject>Chemistry</subject><subject>Chromatography</subject><subject>Chromatography, Liquid - methods</subject><subject>Data interpretation</subject><subject>Digestion</subject><subject>High temperature</subject><subject>Hydrophobicity</subject><subject>Liquid chromatography</subject><subject>Low temperature</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Peptide mapping</subject><subject>Peptide Mapping - methods</subject><subject>Peptides</subject><subject>Peptides - chemistry</subject><subject>Post-translation</subject><subject>Proteolysis</subject><subject>Quality management</subject><subject>Quantitation</subject><subject>Reproducibility of Results</subject><subject>Switching</subject><subject>Tailings</subject><subject>Tandem Mass Spectrometry - methods</subject><subject>Workflow</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU-P0zAQxS0EYsvCN0AoEhcu6Y7t_L0gVdWyi7RVOSziaDn2pPGSxMFOisp5PziO2q2AA6eRNb_3nkePkLcUlhQYvZLKL2UvW9Vgt2QKeMHgGVnQlEGcFQV7ThYAwGOWA1yQV94_AFAKNHtJLnjGk6wEuiCP22E0nfmFOtpM7Wji1Tg6U00jRhscG6ujb9Z9r1v7M1pp7dB70--ijfE-KNYtyr3coY9kH16Ns50c7c7JoTEqupemDfDVWjp3iLd7dJGto9uDdnZobBWILxjCNfrX5EUtW49vTvOSfP10fb--je-2N5_Xq7tYJnk-xgrSkiJTmmYp5xUvEyxqmSW5VlmtK0zrqp7voghFkTIJvC6KGjJdIFRJVfBL8vHoO0xVh1phPzrZisGZTrqDsNKIvze9acTO7kWZpzyEB4MPJwNnf0zoR9EZr7BtZY928oLlIZ8yVvKAvv8HfbCTC33NVJpTllA2GyZHSjnrvcP6_BkKYq5ZhJrFU83iVHOQvfvzkLPoqdcAwBGY5efg_3r-Bk2VumQ</recordid><startdate>20221213</startdate><enddate>20221213</enddate><creator>Kristensen, Dan B.</creator><creator>Ørgaard, Martin</creator><creator>Sloth, Trine M.</creator><creator>Christoffersen, Nanna S.</creator><creator>Leth-Espensen, Katrine Z.</creator><creator>Jensen, Pernille F.</creator><general>American Chemical Society</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>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-5491-1665</orcidid></search><sort><creationdate>20221213</creationdate><title>Optimized Multi-Attribute Method Workflow Addressing Missed Cleavages and Chromatographic Tailing/Carry-Over of Hydrophobic Peptides</title><author>Kristensen, Dan B. ; Ørgaard, Martin ; Sloth, Trine M. ; Christoffersen, Nanna S. ; Leth-Espensen, Katrine Z. ; Jensen, Pernille F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a477t-c0591e2cd16533b394e8fa647dc6fdbe5fbf34691e08852a03f88f06d8e0b4b83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Amino acid sequence</topic><topic>Analytical chemistry</topic><topic>Biopharmaceuticals</topic><topic>Chemistry</topic><topic>Chromatography</topic><topic>Chromatography, Liquid - methods</topic><topic>Data interpretation</topic><topic>Digestion</topic><topic>High temperature</topic><topic>Hydrophobicity</topic><topic>Liquid chromatography</topic><topic>Low temperature</topic><topic>Mass spectrometry</topic><topic>Mass spectroscopy</topic><topic>Peptide mapping</topic><topic>Peptide Mapping - methods</topic><topic>Peptides</topic><topic>Peptides - chemistry</topic><topic>Post-translation</topic><topic>Proteolysis</topic><topic>Quality management</topic><topic>Quantitation</topic><topic>Reproducibility of Results</topic><topic>Switching</topic><topic>Tailings</topic><topic>Tandem Mass Spectrometry - methods</topic><topic>Workflow</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kristensen, Dan B.</creatorcontrib><creatorcontrib>Ørgaard, Martin</creatorcontrib><creatorcontrib>Sloth, Trine M.</creatorcontrib><creatorcontrib>Christoffersen, Nanna S.</creatorcontrib><creatorcontrib>Leth-Espensen, Katrine Z.</creatorcontrib><creatorcontrib>Jensen, Pernille F.</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>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kristensen, Dan B.</au><au>Ørgaard, Martin</au><au>Sloth, Trine M.</au><au>Christoffersen, Nanna S.</au><au>Leth-Espensen, Katrine Z.</au><au>Jensen, Pernille F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Optimized Multi-Attribute Method Workflow Addressing Missed Cleavages and Chromatographic Tailing/Carry-Over of Hydrophobic Peptides</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2022-12-13</date><risdate>2022</risdate><volume>94</volume><issue>49</issue><spage>17195</spage><epage>17204</epage><pages>17195-17204</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Peptide mapping by liquid chromatography mass spectrometry (LC-MS) and the related multi-attribute method (MAM) are well-established analytical tools for verification of the primary structure and mapping/quantitation of co- and post-translational modifications (PTMs) or product quality attributes in biopharmaceutical development. Proteolytic digestion is a key step in peptide mapping workflows, which traditionally is labor-intensive, involving multiple manual steps. Recently, simple high-temperature workflows with automatic digestion were introduced, which facilitate robustness and reproducibility across laboratories. Here, a modified workflow with an automatic digestion step is presented, which includes a two-step digestion at high and low temperatures, as opposed to the original one-step digestion at a high temperature. The new automatic digestion workflow significantly reduces the number of missed cleavages, obtaining a more complete digestion profile. In addition, we describe how chromatographic peak tailing and carry-over is dramatically reduced for hydrophobic peptides by switching from the traditional C18 reversed-phase (RP) column chemistry used for peptide mapping to a less retentive C4 column chemistry. No negative impact is observed on MS/MS-derived sequence coverage when switching to a C4 column chemistry. Overall, the new peptide mapping workflow significantly reduces the number of missed cleavages, yielding more robust and simple data interpretation, while providing dramatically reduced tailing and carry-over of hydrophobic peptides.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>36346901</pmid><doi>10.1021/acs.analchem.2c03820</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0002-5491-1665</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2700 |
| ispartof | Analytical chemistry (Washington), 2022-12, Vol.94 (49), p.17195-17204 |
| issn | 0003-2700 1520-6882 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_9753059 |
| source | MEDLINE; American Chemical Society Journals |
| subjects | Amino acid sequence Analytical chemistry Biopharmaceuticals Chemistry Chromatography Chromatography, Liquid - methods Data interpretation Digestion High temperature Hydrophobicity Liquid chromatography Low temperature Mass spectrometry Mass spectroscopy Peptide mapping Peptide Mapping - methods Peptides Peptides - chemistry Post-translation Proteolysis Quality management Quantitation Reproducibility of Results Switching Tailings Tandem Mass Spectrometry - methods Workflow |
| title | Optimized Multi-Attribute Method Workflow Addressing Missed Cleavages and Chromatographic Tailing/Carry-Over of Hydrophobic Peptides |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-16T13%3A46%3A23IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Optimized%20Multi-Attribute%20Method%20Workflow%20Addressing%20Missed%20Cleavages%20and%20Chromatographic%20Tailing/Carry-Over%20of%20Hydrophobic%20Peptides&rft.jtitle=Analytical%20chemistry%20(Washington)&rft.au=Kristensen,%20Dan%20B.&rft.date=2022-12-13&rft.volume=94&rft.issue=49&rft.spage=17195&rft.epage=17204&rft.pages=17195-17204&rft.issn=0003-2700&rft.eissn=1520-6882&rft_id=info:doi/10.1021/acs.analchem.2c03820&rft_dat=%3Cproquest_pubme%3E2757124129%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2757124129&rft_id=info:pmid/36346901&rfr_iscdi=true |