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

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.

KEYWORDS::

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.

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Order Reprints Request Corporate Permissions

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

Request Academic Permissions

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.

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

Information for

Open access

Opportunities

Help and information

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

Abstract

ACKNOWLEDGMENTS

Reprints and Corporate Permissions

Academic Permissions

Related research

To cite this article:

Download citation

Information for

Open access

Opportunities

Help and information

Keep up to date

Your download is now in progress and you may close this window

Login or register to access this feature