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Detection of Hanganutziu–Deicher antigens in O ‐glycans from pig heart tissues by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry

Detection of Hanganutziu–Deicher antigens in O ‐glycans from pig heart tissues by matrix‐assisted... Background In the α1,3‐galactosyltransferase knockout (α‐GalT KO) pig era, identification of the non‐Gal epitopes is necessary for successful pig‐to‐human xenotransplantation. Recently, we successfully detected α‐Gal epitopes as well as Hanganutziu–Deicher (H‐D) antigens from the N‐glycans in the pig heart tissues, which have been considered as promising non‐Gal antigens. However, the profiling of O‐glycan from pig heart tissues had not been performed owing to the difficulty of O‐glycan preparation. Methods In this study, we established the simple and sensitive method to profile O‐glycans from pig heart aortic valve, aortic wall, pulmonary valve, pulmonary wall, and cardiac muscle tissues. To liberate O‐glycans from the pig heart tissues, we used non‐reductive β‐elimination reagent and subsequently purified the glycans. After permethylation, the glycans were qualitatively analyzed by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS). Results The comprehensive O‐glycan analysis method was successfully validated using model glycoproteins such as bovine serum fetuin (BSF) and bovine submaxillary gland mucin (BSM) glycoproteins, and their O‐glycan profiles were in accordance with the data of previous studies. Next, we applied the method for O‐glycan release and characterization to analysis of various pig heart tissues. As a result, total 39, 33, 24, 36, and 25 of O‐glycans were detected from aortic valve, aortic wall, pulmonary valve, pulmonary wall, and cardiac muscle, respectively. Furthermore, four in aortic valve, one in aortic wall, one in pulmonary valve, one in pulmonary wall, and one in cardiac muscle were particularly determined as terminally N‐glycolylneuraminic acid‐linked O‐glycans, which is considered to be the H‐D antigens. Conclusions Here, we initially described the O‐glycan structures of various pig heart tissues, and additionally, the existence of H‐D antigen type O‐glycans was firstly identified. These results will be fundamental information for overcoming the xenoantigenic carbohydrate‐related immunological rejection in pig‐to‐human heart tissue xenotransplantation. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Xenotransplantation Wiley

Detection of Hanganutziu–Deicher antigens in O ‐glycans from pig heart tissues by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry

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References (24)

Publisher
Wiley
Copyright
© 2013 John Wiley & Sons A/S
ISSN
0908-665X
eISSN
1399-3089
DOI
10.1111/xen.12045
pmid
23855430
Publisher site
See Article on Publisher Site

Abstract

Background In the α1,3‐galactosyltransferase knockout (α‐GalT KO) pig era, identification of the non‐Gal epitopes is necessary for successful pig‐to‐human xenotransplantation. Recently, we successfully detected α‐Gal epitopes as well as Hanganutziu–Deicher (H‐D) antigens from the N‐glycans in the pig heart tissues, which have been considered as promising non‐Gal antigens. However, the profiling of O‐glycan from pig heart tissues had not been performed owing to the difficulty of O‐glycan preparation. Methods In this study, we established the simple and sensitive method to profile O‐glycans from pig heart aortic valve, aortic wall, pulmonary valve, pulmonary wall, and cardiac muscle tissues. To liberate O‐glycans from the pig heart tissues, we used non‐reductive β‐elimination reagent and subsequently purified the glycans. After permethylation, the glycans were qualitatively analyzed by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS). Results The comprehensive O‐glycan analysis method was successfully validated using model glycoproteins such as bovine serum fetuin (BSF) and bovine submaxillary gland mucin (BSM) glycoproteins, and their O‐glycan profiles were in accordance with the data of previous studies. Next, we applied the method for O‐glycan release and characterization to analysis of various pig heart tissues. As a result, total 39, 33, 24, 36, and 25 of O‐glycans were detected from aortic valve, aortic wall, pulmonary valve, pulmonary wall, and cardiac muscle, respectively. Furthermore, four in aortic valve, one in aortic wall, one in pulmonary valve, one in pulmonary wall, and one in cardiac muscle were particularly determined as terminally N‐glycolylneuraminic acid‐linked O‐glycans, which is considered to be the H‐D antigens. Conclusions Here, we initially described the O‐glycan structures of various pig heart tissues, and additionally, the existence of H‐D antigen type O‐glycans was firstly identified. These results will be fundamental information for overcoming the xenoantigenic carbohydrate‐related immunological rejection in pig‐to‐human heart tissue xenotransplantation.

Journal

XenotransplantationWiley

Published: Nov 1, 2013

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