Document Detail


Quantification of fragments of human serum inter-alpha-trypsin inhibitor heavy chain 4 by a surface-enhanced laser desorption/ionization-based immunoassay.
MedLine Citation:
PMID:  16574760     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Several proteolytically derived fragments from the proline-rich region (PRR) of human inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4) have been identified by surface-enhanced or matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS or MALDI-TOF-MS) as potential disease markers. METHODS: Previously, we developed a SELDI-based immunoassay that can simultaneously distinguish and quantify multiple isoforms/variants of a protein/peptide of interest. In this study, we used this high-throughput approach to quantify and characterize the extensive fragmentation within the PRR of human serum ITIH4 and determined its association with different disease conditions. The ITIH4-related fragments were first immunocaptured by use of beads coupled with peptide-specific antibodies. The eluates were then studied by SELDI-TOF-MS. In addition, freshly collected and immediately processed serum and plasma samples were used to analyze the ex vivo stability of these ITIH4 fragments. RESULTS: Human serum ITIH4 was shown to be extensively proteolytically processed within the PRR, and its fragmentation patterns were closely associated with different disease conditions. Fragmentation patterns were generally consistent with cleavages by endoprotease followed by exoprotease actions. Observed fragments changed little under different assay conditions or blood collection and processing procedures. CONCLUSIONS: The fragmentation patterns within the PRR of human serum ITIH4 are associated with different disease conditions and may hold important diagnostic information. These fragmentation patterns could be useful as potential biomarkers for detection and classification of cancer.
Authors:
Jin Song; Manisha Patel; C Nicole Rosenzweig; Yee Chan-Li; Lori J Sokoll; Eric T Fung; Nam-Ho Choi-Miura; Michael Goggins; Daniel W Chan; Zhen Zhang
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2006-03-30
Journal Detail:
Title:  Clinical chemistry     Volume:  52     ISSN:  0009-9147     ISO Abbreviation:  Clin. Chem.     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-05-25     Completed Date:  2006-06-27     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9421549     Medline TA:  Clin Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1045-53     Citation Subset:  IM    
Affiliation:
Center for Biomarker Discovery, Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA.
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MeSH Terms
Descriptor/Qualifier:
Adult
Aged
Amino Acid Sequence
Blood Proteins / chemistry
Breast Neoplasms / blood
Colonic Neoplasms / blood
Diabetes Mellitus / blood
Female
Glycoproteins / blood*,  chemistry
Humans
Immunoassay
Male
Middle Aged
Molecular Sequence Data
Ovarian Neoplasms / blood
Pancreatic Neoplasms / blood
Prostatic Neoplasms / blood
Proteinase Inhibitory Proteins, Secretory
Serum
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
Tumor Markers, Biological / blood
Grant Support
ID/Acronym/Agency:
1P50 CA83639/CA/NCI NIH HHS; CA115102-01/CA/NCI NIH HHS; CA62924/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Blood Proteins; 0/Glycoproteins; 0/ITIH4 protein, human; 0/Proteinase Inhibitory Proteins, Secretory; 0/Tumor Markers, Biological

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


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