Lectin precipitation using phytohemagglutinin-L(4) coupled to avidin-agarose for serological biomarker discovery in colorectal cancer.
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.
Kim
Yong-Sam
YS
Daejeon-KRIBB-FHCRC Research Cooperation Center, KRIBB, 111 Gwahangno, Yuseong-gu, Deajeon, Korea.
Son
Ok Lye
OL
Lee
Ju Yeon
JY
Kim
Sun Hee
SH
Oh
Sejeong
S
Lee
Yoon Suk
YS
Kim
Cheorl-Ho
CH
Yoo
Jong Shin
JS
Lee
Jeong-Hwa
JH
Miyoshi
Eiji
E
Taniguchi
Naoyuki
N
Hanash
Samir M
SM
Yoo
Hyang Sook
HS
Ko
Jeong Heon
JH
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*
D014408:Biomarkers, Tumor; D037102:Lectins; D010835:Phytohemagglutinins; D001360:Avidin; D012685:Sepharose