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
- 2016.3
- 수록면
- 33 - 39 (7page)
이용수
초록· 키워드
오류제보하기
Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.
목차
Abstract
1. INTRODUCTION
2. MATERIALS AND METHODS
3. RESULTS AND DISCUSSION
4. CONCLUSION
REFERENCES
참고문헌 (23)
참고문헌 신청
Browne, S. M. and M. Al-Rubeai (2007) Selection methods for high-producing mammalian cell lines. M. Trend Biotechnol. 9: 425- 432.
Lee, C. J., G. Seth, J. Tsukuda, and R. W. Hamilton (2009) A clone screening method using mRNA levels to determine specific productivity and product quality for monoclonal antibodies. Biotechnolol. Bioeng. 102: 1107-1118.
de la Cruz Edmonds, M. C., M. Tellers, C. Chan, P. Salmon, D. K. Robinson, and J. Markusen (2006) Development of transfection and high-producer screening protocols for the CHOK1SV cell system. Mol. Biotechnol. 34: 179-190.
Fussenegger, M., J. E. Bailey, H. Hauser, and P. P. Mueller (1999) Genetic optimization of recombinant glycoprotein production by mammalian cells. Trends Biotechnol. 17: 35-42.
Wurm, F. M. (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat. Biotechnol. 22: 1393-1398.
Lee, C. J., G. Seth, J. Tsukuda, and R. W. Hamilton (2009) A clone screening method using mRNA levels to determine specific productivity and product quality for monoclonal antibodies. Biotechnolol. Bioeng. 102: 1107-1118.
de la Cruz Edmonds, M. C., M. Tellers, C. Chan, P. Salmon, D. K. Robinson, and J. Markusen (2006) Development of transfection and high-producer screening protocols for the CHOK1SV cell system. Mol. Biotechnol. 34: 179-190.
Fussenegger, M., J. E. Bailey, H. Hauser, and P. P. Mueller (1999) Genetic optimization of recombinant glycoprotein production by mammalian cells. Trends Biotechnol. 17: 35-42.
Wurm, F. M. (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat. Biotechnol. 22: 1393-1398.
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