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제108회 대한화학회 학술발표회, 총회 및 기기전시회 안내 A Simple MS-Approach for Biomarker Discovery and Validation using Non-GlycoPeptide from Glycoproteins in Human Plasma

2011년 8월 1일 21시 10분 03초
BIO1-6 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
목 16시 : 20분
생명화학 - Biomedical Research of Glycoprotein by Mass Spectrometry
저자 및
한국기초과학지원연구원 질량분석연구부, Korea
By targeting Non-glycopeptide which is near glycosylation site in a glycoprotein, we have developed a simple MS approach for biomarker discovery and validation. Because alteration of glycosylation in proteins has been linked to disease such as cancer, glycoproteins have been focused as target biomarkers for early detection, prognosis and therapy of disease. We noticed that proteolytic digestion of glycoprotein could be affected, where enzyme active amino acid is near glycosylation site because of glycan stoichimetric hinderance. It resulted in quantitatively different peptide products according to the degree of glycosylation. In this work, first, this efficiency was proved in comparison using glycosylated AGP(alpha-1 acid glycoprotein standard) digests and deglycosylated AGP digests. Secondly, MS intensities of non-glycopeptides from glycoproteins were relatively higher than those of glycopeptides, thus non-glycopeptides were simply discovered as target biomarkers by label free quantified method without glycoproteins enrichment step. Finally, we confirmed that target non-glycopeptides selected from human plasma analysis were quantitatively different between normal group and disease group by MRM(multiple reaction monitoring) analysis with spiked stable isotope labeled peptide standards. The validation results are accord with the discovery results. Specially, AUC (area under the ROC curve) value of non-glycopeptide from vitronectin precursor protein is excellent as 0.945. Our work simply discovered and validated non-glycopeptide biomarkers from glycoprotein without glycoprotein enrichment by the mass spectrometry (MS)-based proteomics technology.