Drosophila melanogaster S2 cells are more suitable for the production of recombinant COX-1 than Trichoplusia ni BTI TN-5B1-4 cells

  • Chang, Kyung Hwa; 
  • Park, Jong-Hwa; 
  • Hwang-Bo, Jeon; 
  • Chung, Dae Kyun; 
  • Kim, Wonyong; 
  • 외 1명
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초록

Recombinant human cyclooxygenase-1 (COX-1) was expressed from stably transfected Trichoplusia ni BTI TN-5B1-4 (TN-5B1-4) and Drosophila melanogaster S2 cells. Two kinds of recombinant COX-1 with molecular weights (MWs) of 68 and 74 kDa were expressed in the intracellular fractions of stably transfected TN-5B1-4/ COX-1 and S2/COX-1 cells, due to glycosylation. The recombinant COX-1 secreted to medium fractions has a MW of 72 kDa. Recombinant COX-1 in the intracellular fractions was purified to homogeneity using a one-step Ni-NTA affinity fractionation method. Recombinant COX-1 purified from TN-5B1-4/COX-1 and S2/COX-1 cells contained 11,389 and 33,850 Unit/mg of specific peroxidase activity, respectively. The maximum productions of intracellular recombinant COX-1 were 1.7 and 5.6 mu g/10(7) cells in the T-flask cultures of TN-5B1-4/COX-1 and S2/COX-1 cells, respectively. Taken together, our findings indicate that S2 cells can be more suitable system to produce recombinant COX-1, compared to TN-5B1-4 cells.

키워드

Trichoplusia ni BTI TN-5B1-4; Drosophila melanogaster S2; recombinant cyclooxygenase 1; peroxidase activity; PROSTAGLANDIN ENDOPEROXIDE SYNTHASE; BACULOVIRUS EXPRESSION SYSTEM; TN 5B1-4 CELLS; HETEROLOGOUS PROTEINS; CYCLOOXYGENASE 1; UP-REGULATION; INSECT CELLS; LINES; ENDOSTATIN; PROMOTER
제목
Drosophila melanogaster S2 cells are more suitable for the production of recombinant COX-1 than Trichoplusia ni BTI TN-5B1-4 cells
저자
Chang, Kyung Hwa; Park, Jong-Hwa; Hwang-Bo, Jeon; Chung, Dae Kyun; Kim, Wonyong; Chung, In Sik
DOI
10.1007/s12257-014-0239-6
발행일
2014-09
유형
Article
저널명
Biotechnology and Bioprocess Engineering
권
19
호
5
페이지
803 ~ 810