Automated multiplexed affinity-based enrichment of peptides for LC-MS/MS plasma proteomics
Mosquim Junior, Sergio LU and Levander, Fredrik LU
(2024)
In Proteomics
24(21-22).
- Abstract
- Plasma proteomics offers high potential for biomarker discovery, as plasma is collected through a minimally invasive procedure and constitutes the most complex human-derived proteome. However, the wide dynamic range poses a significant challenge. Here, we propose a semi-automated method based on the use of multiple single chain variable fragment antibodies, each enriching for peptides found in up to a few hundred proteins. This approach allows for the analysis of a complementary fraction compared to full proteome analysis. Proteins from pooled plasma were extracted and digested before testing the performance of 29 different antibodies with the aim of reproducibly maximizing peptide enrichment. Our results demonstrate the enrichment of 3662... (More)
- Plasma proteomics offers high potential for biomarker discovery, as plasma is collected through a minimally invasive procedure and constitutes the most complex human-derived proteome. However, the wide dynamic range poses a significant challenge. Here, we propose a semi-automated method based on the use of multiple single chain variable fragment antibodies, each enriching for peptides found in up to a few hundred proteins. This approach allows for the analysis of a complementary fraction compared to full proteome analysis. Proteins from pooled plasma were extracted and digested before testing the performance of 29 different antibodies with the aim of reproducibly maximizing peptide enrichment. Our results demonstrate the enrichment of 3662 peptides not detected in neat plasma or negative controls. Moreover, most antibodies were able to enrich for at least 155 peptides across different levels of abundance in plasma. To further reduce analysis time, a combination of antibodies was used in a multiplexed setting. Repeated sample analyses showed low coefficients of variation, and the method is flexible in terms of affinity binders. It does not impose drastic increases in instrument time, thus showing excellent potential for usage in large scale discovery projects. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/43ed487b-ed54-4583-9ba1-1a00695da7d8
- author
- Mosquim Junior, Sergio
LU
and Levander, Fredrik
LU
- organization
- publishing date
- 2024
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Affinity Enrichment, Automation, Biomarkers, Mass Spectrometry (MS), Plasma Proteomics
- in
- Proteomics
- volume
- 24
- issue
- 21-22
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:39192483
- scopus:85202187168
- ISSN
- 1615-9861
- DOI
- 10.1002/pmic.202400049
- language
- English
- LU publication?
- yes
- id
- 43ed487b-ed54-4583-9ba1-1a00695da7d8
- date added to LUP
- 2024-10-07 15:57:12
- date last changed
- 2025-04-04 14:00:47
@article{43ed487b-ed54-4583-9ba1-1a00695da7d8,
abstract = {{Plasma proteomics offers high potential for biomarker discovery, as plasma is collected through a minimally invasive procedure and constitutes the most complex human-derived proteome. However, the wide dynamic range poses a significant challenge. Here, we propose a semi-automated method based on the use of multiple single chain variable fragment antibodies, each enriching for peptides found in up to a few hundred proteins. This approach allows for the analysis of a complementary fraction compared to full proteome analysis. Proteins from pooled plasma were extracted and digested before testing the performance of 29 different antibodies with the aim of reproducibly maximizing peptide enrichment. Our results demonstrate the enrichment of 3662 peptides not detected in neat plasma or negative controls. Moreover, most antibodies were able to enrich for at least 155 peptides across different levels of abundance in plasma. To further reduce analysis time, a combination of antibodies was used in a multiplexed setting. Repeated sample analyses showed low coefficients of variation, and the method is flexible in terms of affinity binders. It does not impose drastic increases in instrument time, thus showing excellent potential for usage in large scale discovery projects.}},
author = {{Mosquim Junior, Sergio and Levander, Fredrik}},
issn = {{1615-9861}},
keywords = {{Affinity Enrichment; Automation; Biomarkers; Mass Spectrometry (MS); Plasma Proteomics}},
language = {{eng}},
number = {{21-22}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Proteomics}},
title = {{Automated multiplexed affinity-based enrichment of peptides for LC-MS/MS plasma proteomics}},
url = {{http://dx.doi.org/10.1002/pmic.202400049}},
doi = {{10.1002/pmic.202400049}},
volume = {{24}},
year = {{2024}},
}