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Lectin precipitation using phytohemagglutinin‐L 4 coupled to avidin–agarose for serological biomarker discovery in colorectal cancer
Author(s) -
Kim YongSam,
Son Ok Lye,
Lee Ju Yeon,
Kim Sun Hee,
Oh Sejeong,
Lee Yoon Suk,
Kim CheorlHo,
Yoo Jong Shin,
Lee JeongHwa,
Miyoshi Eiji,
Taniguchi Naoyuki,
Hanash Samir M.,
Yoo Hyang Sook,
Ko Jeong Heon
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800034
Subject(s) - lectin , biomarker , biotinylation , microbiology and biotechnology , biomarker discovery , colorectal cancer , chemistry , avidin , cancer , proteomics , agarose , biology , biochemistry , gene , genetics
N ‐acetylglucosaminyltransferase V (GnT‐V) has been reported to be upregulated in malignant cancer cells, and its targets have been sought after with regard to biomarker identification. The low capacity and high false positive rates of 2‐DE gel‐based lectin blots using phytohemagglutinin‐L 4 (L‐PHA) prompted us to develop a novel protocol for identifying GnT‐V targets, in which serum proteins were subjected to immunodepletion, alkylation, and lectin precipitation using L‐PHA coupled to avidin–agarose bead complexes, and tryptic digestion. Proteins captured by L‐PHA conjugates were analyzed by a nano‐LC‐FT‐ICR/LTQ MS. Here, we report 26 candidate biomarkers for colorectal cancer (CRC) that show 100% specificity and sensitivities of greater than 50%. Not only can these candidate proteins be used as analytes for validation, but the novel protocol described herein can be applied to biomarker discovery in nonCRCs.