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Multiplex profiling of tumor‐associated proteolytic activity in serum of colorectal cancer patients
Author(s) -
Yepes Diego,
Costina Victor,
Pilz Lothar R.,
Hofheinz Ralf,
Neumaier Michael,
Findeisen Peter
Publication year - 2014
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201300103
Subject(s) - multiplex , colorectal cancer , protease , medicine , tumor marker , cancer , pathology , oncology , computational biology , biology , bioinformatics , biochemistry , enzyme
Purpose The monitoring of tumor‐associated protease activity in blood specimens has recently been proposed as new diagnostic tool in cancer research. In this paper, we describe the screening of a peptide library for identification of reporter peptides ( RP s) that are selectively cleaved in serum specimens from colorectal cancer patients and investigate the benefits of RP multiplexing. Experimental design A library of 144 RP s was constructed that contained amino acid sequences of abundant plasma proteins. Proteolytic cleavage of RP s was monitored with MS . Five RP s that were selectively cleaved in serum specimens from tumor patients were selected for further validation in serum specimens of colorectal tumor patients ( n = 30) and nonmalignant controls ( n = 60). Results RP spiking and subsequent quantification of proteolytic fragments with LC‐MS showed good reproducibility with CV s always below 26%. The linear discriminant analysis and PCA revealed that a combination of RP s for diagnostic classification is superior to single markers. Classification accuracy reached 88% (79/90) when all five markers were combined. Conclusions and clinical relevance Functional protease profiling with RP s might improve the laboratory‐based diagnosis, monitoring and prognosis of malignant disease, and has to be evaluated thoroughly in future studies.