KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Serum Free Medium Development for Recombinant Erythropoietin Production using Novel Cell Line (QT35)

QT35 세포주에서 제조합 에리스로포이에틴 생산을 위한 무혈청 배지의 개발

  • 주형민 (서울대학교 유전공학연구소 및 서울대학교 공과대학 응용화학부) ;
  • 김병기 (서울대학교 유전공학연구소 및 서울대학교 공과대학 응용화학부) ;
  • 김선영 (서울대학교 유전공학연구소) ;
  • 김태한 ((주) 일동제약 중앙연구소) ;
  • 김태용 ((주) 일동제약 중앙연구소)
  • Published : 1998.06.01
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Abstract

Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.

Keywords

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