Document Detail


Investigation and reduction of sub-microgram peptide loss using molecular weight cut-off fractionation prior to mass spectrometric analysis.
MedLine Citation:
PMID:  23019164     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This work investigates the introduction of methanol and a salt modifier to molecular weight cut-off membrane-based centrifugal filters (MWCO) to enrich sub-microgram peptide quantities. Using a neuropeptide standard, bradykinin, sample loss was reduced over two orders of magnitude with and without undigested protein present. Additionally, a bovine serum albumin tryptic digestion was investigated. Twenty-seven tryptic peptides were identified from MALDI mass spectra after enriching with methanol while only two tryptic peptides were identified after MWCO separation using H(2)O. The strategy presented here enhances recovery from MWCO separation for sub-µg peptide samples.
Authors:
Robert Cunningham; Jingxin Wang; Daniel Wellner; Lingjun Li
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Journal of mass spectrometry : JMS     Volume:  47     ISSN:  1096-9888     ISO Abbreviation:  J Mass Spectrom     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-09-28     Completed Date:  2013-01-23     Revised Date:  2013-07-31    
Medline Journal Info:
Nlm Unique ID:  9504818     Medline TA:  J Mass Spectrom     Country:  England    
Other Details:
Languages:  eng     Pagination:  1327-32     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 John Wiley & Sons, Ltd.
Affiliation:
Department of Chemistry and School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bradykinin / analysis,  chemistry
Cattle
Humans
Methanol / chemistry
Molecular Weight
Peptide Fragments / analysis,  chemistry
Peptides / analysis*,  chemistry
Serum Albumin, Bovine / analysis,  chemistry
Sodium Chloride / chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Trypsin / chemistry
Water / chemistry
Grant Support
ID/Acronym/Agency:
P50 AG033514/AG/NIA NIH HHS
Chemical
Reg. No./Substance:
0/Peptide Fragments; 0/Peptides; 0/Serum Albumin, Bovine; 58-82-2/Bradykinin; 67-56-1/Methanol; 7647-14-5/Sodium Chloride; 7732-18-5/Water; EC 3.4.21.4/Trypsin
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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