지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2017.9
- 수록면
- 193 - 198 (6page)
- DOI
- 10.7841/ksbbj.2017.32.3.193
이용수
초록· 키워드
오류제보하기
Sialylation is important in producing therapeutic proteins such as antibody, cytokine and fusion protein. Thus, enhancement of sialylation is usually performed in CHO cell cultures. α2,6-Sialyltransferase (ST), which plays a key role in the attachment of α2,6-sialic acid, is present in human cells but not in Chinese hamster ovary (CHO) cells. Overexpression of α2,6-ST can be used for enhancing the degree of sialylation and achieving human-like glycosylation. In this study, we constructed CHO cells producing human cytotoxic T-lymphocyte antigen4-immunoglobulin (hCTLA4-Ig) as well as α2,6- ST. Transfected CHO cells were selected using G418 and stable cell line was established. Profiles of viable cell density and hCTLA4-Ig titer in an overexpressed cell line were similar to those of a wild-type cell line. It was confirmed that the total amount of sialic acid was increased and α2,6-sialic acid was attached to the terminal residues of N-glycan of hCTLA4-Ig by ESI-LC-MS. Compared to 100% of α2,3-sialic acid in wild type cells, 70.9% of total sialylated N-glycans were composed of α2,6-sialic acid in transfected cells. In conclusion, overexpression of α2,6-ST in CHO cells led to the increase of both the amount of total sialylated N-glycan and the content of α2,6- sialic acid, which is more resemble to human-like structure of glycosylation.
목차
Abstract
1. INTRODUCTION
2. MATERIALS AND METHODS
3. RESULTS AND DISCUSSION
4. CONCLUSION
REFERENCES
참고문헌 (17)
참고문헌 신청
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Mullard, A. (2017) 2016 FDA drug approvals. Nat. Rev. Drug Discov. 16: 73-76.
Lalonde, M. E. and Y. Durocher (2017) Therapeutic glycoprotein production in mammalian cells. J. Biotechnol. 251: 128-140.
Xu, X., H. Nagarajan, N. E. Lewis, S. Pan, Z. Cai, X. Liu, W. Chen, M. Xie, W. Wang, S. Hammond, M. R. Andersen, N. Neff, B. Passarelli, W. Koh, H. C. Fan, J. Wang, Y. Gui, K. H. Lee, M. J. Betenbaugh, S. R. Quake, I. Famili, B. O. Palsson, and J. Wang (2011) The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line. Nat. Biotechnol. 29:735-741.
Bosques, C. J., B. E. Collins, J. W. Meador, H. Sarvaiya, J. L. Murphy, G. Dellorusso, D. A. Bulik, I. -S. Hsu, N. Washburn, S. F. Sipsey, J. R. Myette, R. Raman, Z. Shriver, R. Sasisekharan, and G. Venkataraman (2010) Chinese hamster ovary cells can produce galactose-a-1,3-galactose antigens on proteins. Nat. Biotechnol. 28: 1153-1156.
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