High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device
15 páginas, 8 figuras, 2 tablas
| Autores: | , , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2026 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/422988 |
| Acceso en línea: | http://hdl.handle.net/10261/422988 |
| Access Level: | acceso abierto |
| Palabra clave: | PAC SP3 High-throughput sample preparation Oasis HLB Phosphoproteomics Proteomics |
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High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin DeviceRoumelioti, GeorgiaMontoya, AlexFisher, Gemma L. M.Pascual Navarro, EnekoWoods, AngelaBennett, JaneNavaratnam, NaveenanGonzalez-Carvajal, OliverBirch, JodiePyman, ElizabethYu, SijiaGruevska, AleksandraVuillemenot, Luc-AlbanLushchak, OlehHall, ZoeBarr, Alexis R.Speck, ChristianVernia, SantiagoScott, William R.Gil, JesusAragon, LuisFets, LouiseCarling, DavidShliaha, Pavel V.PACSP3High-throughput sample preparationOasis HLBPhosphoproteomicsProteomics15 páginas, 8 figuras, 2 tablasHigh-throughput proteomics requires efficient and highly reproducible sample processing, yet workflows─particularly for PTM profiling─remain complex and costly to fully automate. Here, we present a practical intermediate solution using manually operated 96-channel devices: the Gilson Platemaster P220 pipettor and VP Scientific 96-well magnetic pin device. Using this setup, we achieved robust and reproducible phosphoproteomics in a 96-well format, completing protein aggregation capture (PAC/SP3) digestion, desalting, phosphopeptide enrichment, and a second desalting step within 2 days while minimizing operator workload and variability. Several innovations enable this workflow. First, we describe a cost-efficient method to generate 96-well solid-phase extraction plates by directly packing the Oasis HLB sorbent into tapered filter plates. We extensively characterize these plates in terms of loading capacity, lipid removal efficiency, and suitability for high-pH fractionation. Second, we demonstrate that efficient PAC digestion does not require continuous bead suspension; instead, digestion can be achieved by briefly aspirating beads in protease solution, eliminating the need for orbital shaking and simplifying automation. The presented workflow familiarizes users with 96-channel devices and hence serves as a good step toward full automation.Work at the proteomics core facility is supported by MRC LMS core funding (MC-A651-53297).Peer reviewedAmerican Chemical SocietyMRC Laboratory of Medical SciencesVernia, Santiago [0000-0001-6728-5555]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202620262026info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/422988reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.1021/acs.jproteome.5c01020Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4229882026-05-22T06:33:51Z |
| dc.title.none.fl_str_mv |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| title |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| spellingShingle |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device Roumelioti, Georgia PAC SP3 High-throughput sample preparation Oasis HLB Phosphoproteomics Proteomics |
| title_short |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| title_full |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| title_fullStr |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| title_full_unstemmed |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| title_sort |
High-Throughput Proteomics Sample Preparation Using a 96-Channel Pipettor and Magnetic Pin Device |
| dc.creator.none.fl_str_mv |
Roumelioti, Georgia Montoya, Alex Fisher, Gemma L. M. Pascual Navarro, Eneko Woods, Angela Bennett, Jane Navaratnam, Naveenan Gonzalez-Carvajal, Oliver Birch, Jodie Pyman, Elizabeth Yu, Sijia Gruevska, Aleksandra Vuillemenot, Luc-Alban Lushchak, Oleh Hall, Zoe Barr, Alexis R. Speck, Christian Vernia, Santiago Scott, William R. Gil, Jesus Aragon, Luis Fets, Louise Carling, David Shliaha, Pavel V. |
| author |
Roumelioti, Georgia |
| author_facet |
Roumelioti, Georgia Montoya, Alex Fisher, Gemma L. M. Pascual Navarro, Eneko Woods, Angela Bennett, Jane Navaratnam, Naveenan Gonzalez-Carvajal, Oliver Birch, Jodie Pyman, Elizabeth Yu, Sijia Gruevska, Aleksandra Vuillemenot, Luc-Alban Lushchak, Oleh Hall, Zoe Barr, Alexis R. Speck, Christian Vernia, Santiago Scott, William R. Gil, Jesus Aragon, Luis Fets, Louise Carling, David Shliaha, Pavel V. |
| author_role |
author |
| author2 |
Montoya, Alex Fisher, Gemma L. M. Pascual Navarro, Eneko Woods, Angela Bennett, Jane Navaratnam, Naveenan Gonzalez-Carvajal, Oliver Birch, Jodie Pyman, Elizabeth Yu, Sijia Gruevska, Aleksandra Vuillemenot, Luc-Alban Lushchak, Oleh Hall, Zoe Barr, Alexis R. Speck, Christian Vernia, Santiago Scott, William R. Gil, Jesus Aragon, Luis Fets, Louise Carling, David Shliaha, Pavel V. |
| author2_role |
author author author author author author author author author author author author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
MRC Laboratory of Medical Sciences Vernia, Santiago [0000-0001-6728-5555] Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72] |
| dc.subject.none.fl_str_mv |
PAC SP3 High-throughput sample preparation Oasis HLB Phosphoproteomics Proteomics |
| topic |
PAC SP3 High-throughput sample preparation Oasis HLB Phosphoproteomics Proteomics |
| description |
15 páginas, 8 figuras, 2 tablas |
| publishDate |
2026 |
| dc.date.none.fl_str_mv |
2026 2026 2026 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article http://purl.org/coar/resource_type/c_6501 Publisher's version info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10261/422988 |
| url |
http://hdl.handle.net/10261/422988 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
https://doi.org/10.1021/acs.jproteome.5c01020 Sí |
| dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess |
| eu_rights_str_mv |
openAccess |
| dc.publisher.none.fl_str_mv |
American Chemical Society |
| publisher.none.fl_str_mv |
American Chemical Society |
| dc.source.none.fl_str_mv |
reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC instname:Consejo Superior de Investigaciones Científicas (CSIC) |
| instname_str |
Consejo Superior de Investigaciones Científicas (CSIC) |
| reponame_str |
DIGITAL.CSIC. Repositorio Institucional del CSIC |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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|
| repository.mail.fl_str_mv |
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1869419719090503680 |
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15,812455 |