title: Lectin precipitation using phytohemagglutinin-L(4) coupled to avidin-agarose for serological biomarker discovery in colorectal cancer.
abstract: 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.
authors.affiliation: Daejeon-KRIBB-FHCRC Research Cooperation Center, KRIBB, 111 Gwahangno, Yuseong-gu, Deajeon, Korea.
authors.forename: Yong-Sam
authors.initials: YS
authors.lastname: Kim
authors.forename: Ok Lye
authors.initials: OL
authors.lastname: Son
authors.forename: Ju Yeon
authors.initials: JY
authors.lastname: Lee
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Kim
authors.forename: Sejeong
authors.initials: S
authors.lastname: Oh
authors.forename: Yoon Suk
authors.initials: YS
authors.lastname: Lee
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
authors.forename: Jong Shin
authors.initials: JS
authors.lastname: Yoo
authors.forename: Jeong-Hwa
authors.initials: JH
authors.lastname: Lee
authors.forename: Eiji
authors.initials: E
authors.lastname: Miyoshi
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
authors.forename: Samir M
authors.initials: SM
authors.lastname: Hanash
authors.forename: Hyang Sook
authors.initials: HS
authors.lastname: Yoo
authors.forename: Jeong Heon
authors.initials: JH
authors.lastname: Ko
mesh_terms: D001360:Avidin / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000097:blood* / Q000737:chemistry; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D015179:Colorectal Neoplasms / Q000097:blood*; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D013058:Mass Spectrometry; D010835:Phytohemagglutinins / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry*
chemical_list: D014408:Biomarkers, Tumor; D037102:Lectins; D010835:Phytohemagglutinins; D001360:Avidin; D012685:Sepharose