Preparative Release and Mass Spectrometric Analysis of Tn and T Antigens Existing in Sericin

doi:10.7503/cjcu20230378

Abstract

Abstract:

Silkworm sericin-derived Tn and T antigens， a group of biomarkers of many cancers， were targeted in this study. The O-glycans of sericin were first released using the “one-pot” non-reductive release method， the ammonia-catalyzed non-reductive release strategy and the pronase E digestion approach. The samples were subsequently fractionated for purification using solid-phase extraction columns and separated for preparation via high-performance liquid chromatography（HPLC）. The generated glycan samples were finally analyzed in detail by means of electrospray ionization mass spectrometry（ESI-MS）， tandem mass spectrometry（MS/MS） and liquid chromatography coupling online with mass spectrometry（Online LC-MS）. The obtained results showed that the O-glycans of sericin just include O-GalNAc and O-GalNAcGal， the contents of which have been determined to be 1.58 μg/g and 0.54 μg/g， respectively. Their molar ratio in sericin is approximately 5.3∶1. The O-linked monosaccharide of sericin， GalNAc， allows efficient non-reductive release as a reducing form in ammonium hydroxide and fine separation by liquid chromatography. Tn and T antigens can be efficiently generated from sericin using pronase E with an amount equal to the raw glycoprotein.

Key words: Tn and T antigens, Sericin, O-Glycan release, High-performance liquid chromatography, Mass spectrometry

CLC Number:

O629.1

TrendMD:

MAN Lijuan, YE Jiajia, ZHANG Defeng, ZOU Zihua, HUANG Linjuan, WANG Zhongfu, WANG Chengjian. Preparative Release and Mass Spectrometric Analysis of Tn and T Antigens Existing in Sericin[J]. Chem. J. Chinese Universities, 2023, 44(12): 20230378.

Figures/Tables 4

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