KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Engineering Human-like Sialylation in CHO Cells Producing hCTLA4-Ig by Overexpressing α2,6-Sialyltransferase

α2,6-Sialyltransferase 과발현을 통한 인간형 시알산 부가 hCTLA4-Ig 생산 CHO 세포주 제작

  • 임진혁 (인하대학교 공과대학 생물공학과) ;
  • 차현명 (인하대학교 공과대학 생물공학과) ;
  • 박혜진 (중앙대학교 약학대학 약학부) ;
  • 김하형 (중앙대학교 약학대학 약학부) ;
  • 김동일 (인하대학교 공과대학 생물공학과)
  • Received : 2017.07.12
  • Accepted : 2017.08.21
  • Published : 2017.09.30
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Abstract

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. ${\alpha}2,6$-Sialyltransferase (ST), which plays a key role in the attachment of ${\alpha}2,6-sialic$ acid, is present in human cells but not in Chinese hamster ovary (CHO) cells. Overexpression of ${\alpha}2,6-ST$ 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 ${\alpha}2,6-ST$. 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 ${\alpha}2,6-sialic$ acid was attached to the terminal residues of N-glycan of hCTLA4-Ig by ESI-LC-MS. Compared to 100% of ${\alpha}2,3-sialic$ acid in wild type cells, 70.9% of total sialylated N-glycans were composed of ${\alpha}2,6-sialic$ acid in transfected cells. In conclusion, overexpression of ${\alpha}2,6-ST$ in CHO cells led to the increase of both the amount of total sialylated N-glycan and the content of ${\alpha}2,6-sialic$ acid, which is more resemble to human-like structure of glycosylation.

Keywords

Acknowledgement

Supported by : 한국연구재단

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