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Current Eye Research Volume 34, 2009 - Issue 10
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Original Article

Identification of α-Gal and non-Gal Epitopes in Pig Corneal Endothelial Cells and Keratocytes by Using Mass Spectrometry

Yun-Gon KimInstitute of Molecular Biology and Genetics and Institute of Bioengineering, Seoul National University, Seoul, Korea
,
Joo Youn OhDepartment of Ophthalmology, Seoul Artificial Eye Center, and Xenotransplantation Research Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea
,
Geun-Cheol GilSchool of Chemical and Biological Engineering, Seoul National University, Seoul, Korea
,
Mee Kum KimDepartment of Ophthalmology, Seoul National University College of Medicine, and Seoul Artificial Eye Center and Xenotransplantation Research Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea
,
Jung Hwa KoDepartment of Ophthalmology, Seoul National University College of Medicine, and Seoul Artificial Eye Center and Xenotransplantation Research Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea
,
Sukmook LeeCancer Research Institute, Seoul National University College of Medicine, Seoul, Korea
,
Hyun Ju LeeDepartment of Ophthalmology, Seoul National University College of Medicine, and Seoul Artificial Eye Center and Xenotransplantation Research Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea
,
Won Ryang WeeDepartment of Ophthalmology, Seoul National University College of Medicine and Seoul Artificial Eye Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea
&
Byung-Gee KimInstitute of Molecular Biology and Genetics, School of Chemical and Biological Engineering, and Institute of Bioengineering, Seoul National University, Seoul, KoreaCorrespondence[email protected]
 show all
Pages 877-895 | Received 22 Oct 2008, Accepted 13 Jul 2009, Published online: 06 Nov 2009
 
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Abstract

Purpose: To investigate the expression of α-Gal or unidentified non-Gal antigens in pig corneal endothelial cells and keratocytes, we performed the qualitative and quantitative analysis by using mass spectrometry.

Methods: The N-glycans from common adult pig corneal endothelial cells and keratocytes cultured in vitro were directly analyzed by using mass spectrometric approaches. In addition, immunochemical staining was added to confirm the non-Gal antigen expression in pig corneal cells.

Results: Totally, 34 of the sialylated N-glycans from pig corneal endothelial cells and 27 from pig keratocytes were identified and observed to contain nonhuman sialic acid, NeuGc as well as NeuAc. In addition, we were able to detect 25 of α-galactosylated N-glycan structures (22.2% of total) from the pig corneal endothelial cells and 18 of that (17.5% of total) from the pig keratocytes by using mass spectrometric approaches. On immunofluorescent staining, the expression of sialylated glycans was also observed.

Conclusions: As well as α-Gal epitopes, several promising non-Gal antigens were widely expressed on both pig corneal endothelial cells and keratocytes. The detailed structural information of the α-Gal and non-Gal epitopes would be a tremendous value to develop a new strategy for the successful corneal xenotransplantation in future.

ACKNOWLEDGMENTS

This work was supported by a grant (M10417060004-04N1706-00410) from Korea Biotech R&D Group of the Next-Generation Growth Engine Project of the Ministry of Education, Science and Technology, and by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MOST) (No. R0A-2007-000-10007-0). This study was also supported by grant of the Korea health 21 RND project, the ministry and welfare, Republic of Korea. (Project No. A04-0004-AZ1205-06A3-00100B).

Declaration of interest: The authors report no conflict of interest. The authors alone are responsible for the content and writing of the paper.

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