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Adding polyvinylpyrrolidone to low level protein samples significantly improves peptide recovery in FASP digests: an inexpensive and simple modification to the FASP protocol

From National Research Council Canada

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DOIResolve DOI: https://doi.org/10.1016/j.jprot.2020.104000
AuthorSearch for: 1; Search for: 1
Affiliation
  1. National Research Council Canada. Human Health Therapeutics
FunderSearch for: National Research Council Canada. Human Health Therapeutics Research Centre
FormatText, Article
Physical description5 p.
Subjectfilter-aided sample preparation; polyvinylpyrrolidone; proteomics; sample preparation
Abstract
Filter-aided sample preparation (FASP) remains a popular choice for proteomic sample preparation, particularly for its ability to produce a ‘clean’ peptide sample clear of large molecule contaminants. However, sample loss continues to be a problem particularly for sample inputs that contain less than ten micrograms of protein. Here, we describe that the simple addition of a polymer, polyvinylpyrrolidone-40 (PVP-40) to the protein sample prior to FASP digest significantly improves peptide recovery and identifications, especially with lower level sample inputs. PVP-FASP produces clean samples which required no additional sample clean-up prior to nanoLC-MS analysis. In addition, PVP-FASP is compatible with other FASP modifications, including the use of sodium deoxycholate (DOC) to improve trypsin digestion.
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PublisherElsevier
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  • Journal of Proteomics 230, no. C, 104000 (6 January 2021).
LanguageEnglish
Peer reviewedYes
IdentifierS1874391920303687
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Record identifier9db0930d-2b17-4e68-89c9-46454856dc5a
Record created2022-09-02
Record modified2025-12-18
Date modified: