Development of an all-in-one technology for glycan profiling targeting formalin-embedded tissue sections |
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Authors: | Matsuda Atsushi Kuno Atsushi Ishida Hiroyasu Kawamoto Toru Shoda Jun-Ichi Hirabayashi Jun |
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Institution: | a Lectin Application and Analysis Team, Research Center for Medical Glycoscience (RCMG), National Institute of Advanced Industrial Science and Technology (AIST), AIST Tsukuba Central 2, 1-1-1 Umezono, Tsukuba-shi, Ibaraki 305-8568, Japan b Department of Gastroenterology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba-shi, Ibaraki 305-8577, Japan c Mito Medical Center, Ibaraki-cho, Higashiibarakigun, Ibaraki 311-3193, Japan d Gasterointestinal Surgery, Sojinkai Hospital, Toride-shi, Ibaraki 300-1546, Japan |
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Abstract: | An ultra-sensitive method for glycan analysis targeting small tissue sections (1.5 mm in diameter) is described as an application of a recently-established lectin microarray technology. The developed system achieved a high level of detection of a tissue section consisting of approximately 500 cells for differential profiling, where both N- and O-glycans attached to a pool of glycoproteins are subjected to multiplex analysis with 43 lectins. By using an optimized protocol for differential glycan analysis, sections of adenocarcinoma (n = 28) and normal epithelia (n = 12) of the colon were analyzed in an all-in-one manner. As a result, Wisteria floribunda agglutinin (WFA) was found to clearly differentiate cancerous from normal epithelia with P < 0.0001. The obtained results correlated well with the subsequent histochemical study using biotinylated WFA. Thus, the developed technology proved to be valid for expanding the lectin microarray applications to tissue-based glycomics, and hence, should accelerate a discovery phase of glycan-related biomarkers. |
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Keywords: | Gal galactose GalNAc N-acetylgalactosamine GlcNAc N-acetylglucosamine MUC mucin core protein SE succimidyl ester Sia sialic acid PBS 10 mM phosphate-buffered saline pH 7 0 TBS Tris-buffered saline pH 7 4 DAB 3 3&prime -diaminobenzidine BSA bovine serum albumin NP-40 Nonidet P-40 |
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