Document Detail


Systematic discovery of ectopic pregnancy serum biomarkers using 3-D protein profiling coupled with label-free quantitation.
MedLine Citation:
PMID:  21142075     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Ectopic pregnancy (EP) and normal intrauterine pregnancy (IUP) serum proteomes were quantitatively compared to systematically identify candidate biomarkers. A 3-D biomarker discovery strategy consisting of abundant protein immunodepletion, SDS gels, LC-MS/MS, and label-free quantitation of MS signal intensities identified 70 candidate biomarkers with differences between groups greater than 2.5-fold. Further statistical analyses of peptide quantities were used to select the most promising 12 biomarkers for further study, which included known EP biomarkers, novel EP biomarkers (ADAM12 and ISM2), and five specific isoforms of the pregnancy specific beta-1-glycoprotein family. Technical replicates showed good reproducibility and protein intensities from the label-free discovery analysis compared favorably with reported abundance levels of several known reference serum proteins over at least 3 orders of magnitude. Similarly, relative abundances of candidate biomarkers from the label-free discovery analysis were consistent with relative abundances from pilot validation assays performed for five of the 12 most promising biomarkers using label-free multiple reaction monitoring of both the patient serum pools used for discovery and the individual samples that constituted these pools. These results demonstrate robust, reproducible, in-depth 3-D serum proteome discovery, and subsequent pilot-scale validation studies can be achieved readily using label-free quantitation strategies.
Authors:
Lynn A Beer; Hsin-Yao Tang; Sira Sriswasdi; Kurt T Barnhart; David W Speicher
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-01-07
Journal Detail:
Title:  Journal of proteome research     Volume:  10     ISSN:  1535-3907     ISO Abbreviation:  J. Proteome Res.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-03-04     Completed Date:  2011-06-13     Revised Date:  2014-10-15    
Medline Journal Info:
Nlm Unique ID:  101128775     Medline TA:  J Proteome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1126-38     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
ADAM Proteins / blood,  genetics
Amino Acid Sequence
Biological Markers / blood*
Blood Proteins / analysis*
Chromatography, Liquid / methods
Databases, Protein
Electrophoresis, Polyacrylamide Gel / methods
Female
Humans
Membrane Proteins / blood,  genetics
Molecular Sequence Data
Pregnancy / blood*
Pregnancy, Ectopic / blood*
Proteome / analysis*
Proteomics / methods*
Tandem Mass Spectrometry / methods
Grant Support
ID/Acronym/Agency:
CA10815/CA/NCI NIH HHS; HD036455/HD/NICHD NIH HHS; K24 HD060687/HD/NICHD NIH HHS; R01 HD036455/HD/NICHD NIH HHS; R01 HD036455-10/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Blood Proteins; 0/Membrane Proteins; 0/Proteome; EC 3.4.24.-/ADAM 12 protein; EC 3.4.24.-/ADAM Proteins
Comments/Corrections

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


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