Close
Help
Need Help?





JOURNAL

Biomarker Insights

778,817 Journal Article Views | Journal Analytics

Endogenous Plasma Peptide Detection and Identification in the Rat by a Combination of Fractionation Methods and Mass Spectrometry

Submit a Paper



Publication Date: 09 Oct 2007

Journal: Biomarker Insights

Citation: Biomarker Insights 2007:2 385-401

Fabrice Bertile, Flavie Robert, Véronique Delval-Dubois, Sarah Sanglier, Christine Schaeffer and Alain Van Dorsselaer

Institut Pluridisciplinaire Hubert Curien, Département Sciences Analytiques, Laboratoire de Spectrométrie de Masse Bio-Organique, CNRS-ULP UMR 7178, ECPM, 25 rue Becquerel, 67087 Strasbourg Cedex 2, France.

Abstract: Mass spectrometry-based analyses are essential tools in the field of biomarker research. However, detection and characterization of plasma low abundance and/or low molecular weight peptides is challenged by the presence of highly abundant proteins, salts and lipids. Numerous strategies have already been tested to reduce the complexity of plasma samples. The aim of this study was to enrich the low molecular weight fraction of rat plasma. To this end, we developed and compared simple protocols based on membrane filtration, solid phase extraction, and a combination of both. As assessed by UV absorbance, an albumin depletion 99% was obtained. The multistep fractionation strategy (including reverse phase HPLC) allowed detection, in a reproducible manner (CV [1] 30%–35%), of more than 450 peaks below 3000 Da by MALDI-TOF/MS. A MALDI-TOF/MS-determined LOD as low as 1 fmol/μL was obtained, thus allowing nanoLC-Chip/ MS/MS identification of spiked peptides representing ∼10–6% of total proteins, by weight. Signal peptide recovery ranged between 5%–100% according to the spiked peptide considered. Tens of peptide sequence tags from endogenous plasma peptides were also obtained and high confidence identifications of low abundance fibrinopeptide A and B are reported here to show the efficiency of the protocol. It is concluded that the fractionation protocol presented would be of particular interest for future differential (high throughput) analyses of the plasma low molecular weight fraction.


Downloads

PDF  (553.61 KB PDF FORMAT)

RIS citation   (ENDNOTE, REFERENCE MANAGER, PROCITE, REFWORKS)

BibTex citation   (BIBDESK, LATEX)

XML

PMC HTML


Sharing


Our Service Promise

  • Prompt Processing (3 Weeks to Editorial Decision)
  • Fair, Independent Peer Review
  • High Visibility & Extensive Indexing
What Your Colleagues Say About Biomarker Insights
testimonial_image
I have had a great experience with submitting my cancer prognosis study to Biomarker Insights. The comments from reviewers and associate editor are high quality and insightful. Congratulations and keep up the good work.
Dr Yi Hong Zhou (University of California, Irvine, CA, USA)
More Testimonials

Quick Links




Follow Us We make it easy to find new research papers.
Email Alerts RSS Feeds
Facebook Google+ Twitter
Pinterest Tumblr YouTube




SUBJECT HUBS
Author Survey Results
author_survey_results
All authors are surveyed after their articles are published. Authors are asked to rate their experience in a variety of areas, and their responses help us to monitor our performance. Presented here are their responses in some key areas. No 'poor' or 'very poor' responses were received; these are represented in the 'other' category.
See Our Results