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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제23권 제8호
- 발행연도
- 2013.1
- 수록면
- 1,176 - 1,184 (9page)
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Anti-hepatocyte growth factor (anti-HGF) monoclonal antibodies (mAbs) are potential therapeutics against various cancers. Screening for high-producer clones is a time-consuming and complex process and is a major hurdle in the development of therapeutic mAbs. Here, we describe an efficient approach that allows the selection of high-producer Chinese hamster ovary (CHO) cell lines producing the novel anti-HGF mAb SFN68, which was generated previously by immunizing HGF bound to its receptor c-Met. We selected an SFN68-producing parental cell line via transfection of the dihydrofolate reductase-deficient CHO cell line DG44,which was preadapted to serum-free suspension culture, with an SFN68-expression vector.
Subsequent gene amplification via multiple passages of the parental cell line in a methotrexate-containing medium over 4 weeks, followed by clonal isolation, enabled us to isolate two cell lines, 2F7 and 2H4, with 3-fold higher specific productivity. We also screened 72 different media formulated with diverse feed and basal media to develop a suboptimized medium. In the established suboptimized medium, the highest anti-HGF mAb yields of the 2F7 and 2H4 clones were 842 and 861 mg/l, respectively, which were about 10.5-fold higher than that of the parental cell line in a non-optimized basal medium. The selected CHO cell lines secreting high titers of SFN68 would be useful for the production of sufficient amounts of antibodies for efficacy evaluation in preclinical and early clinical studies.
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참고문헌 (22)
참고문헌 신청
Backliwal G / 2008 / Rational vector design and multipathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g/l by transient transfection under se
Bottaro DP / 1991 / Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product / Science 251 : 802 ~ 804
Burgess T / 2006 / Fully human monoclonal antibodies to hepatocyte growth factor with therapeutic potential against hepatocyte growth factor/c-Met-dependent human tumors / Ca
Cacciatore JJ / 2010 / Gene amplification and vector engineering to achieve rapid and high-level therapeutic protein production using the Dhfr-based CHO cell selection system
Cao B / 2001 / Neutralizing monoclonal antibodies to hepatocyte growth factor/scatter factor (HGF/SF) display antitumor activity in animal models / Proc. Natl. Acad. Sci. USA
Bottaro DP / 1991 / Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product / Science 251 : 802 ~ 804
Burgess T / 2006 / Fully human monoclonal antibodies to hepatocyte growth factor with therapeutic potential against hepatocyte growth factor/c-Met-dependent human tumors / Ca
Cacciatore JJ / 2010 / Gene amplification and vector engineering to achieve rapid and high-level therapeutic protein production using the Dhfr-based CHO cell selection system
Cao B / 2001 / Neutralizing monoclonal antibodies to hepatocyte growth factor/scatter factor (HGF/SF) display antitumor activity in animal models / Proc. Natl. Acad. Sci. USA
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