Overproduction of recombinant human transforming growth factor beta 3 in Chinese hamster ovary cells.
Transforming growth factor beta 3 (TGFβ3) is an important cytokine, functioning in cell proliferation and differentiation, and has been considered to have therapeutic potential for treating various diseases and for scar reduction in adult wound healing. In the current study, a Chinese hamster ovary (CHO) cell line overexpressing recombinant human TGFβ3 (rhTGFβ3) was established. Through a 15-day fed-batch culture process in a 7.5-l bioreactor (5-l working volume) using chemically defined medium, the established cells could produce over 133mg/l of rhTGFβ3 protein. The rhTGFβ3 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 65%, with protein purity greater than 97%. The N-terminal amino acid sequences of the purified rhTGFβ3 were confirmed by N-terminal sequencing analysis. The purified rhTGFβ3 was further demonstrated to be functionally active by measuring the inhibition of growth of HT-2 cells, revealing a half-maximal effective concentration of 42.11pg/ml and specific activity of 1.84×10(7)U/mg.
Graduate School of Biotechnology, Kyung Hee University, 1732 Deogyeong-daero, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
Byungseok
B
Choi
Graduate School of Biotechnology, Kyung Hee University, 1732 Deogyeong-daero, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
Yujin
Y
Lee
Graduate School of Biotechnology, Kyung Hee University, 1732 Deogyeong-daero, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
Jia
J
Pi
Graduate School of Biotechnology, Kyung Hee University, 1732 Deogyeong-daero, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
Yongsu
Y
Jeong
Graduate School of Biotechnology, Kyung Hee University, 1732 Deogyeong-daero, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
Kwangehee
K
Baek
Graduate School of Biotechnology, Kyung Hee University, 1732 Deogyeong-daero, Yongin-si, Gyeonggi-do 446-701, Republic of Korea. Electronic address: jsyoon@khu.ac.kr.
Jaeseung
J
Yoon
D000595:Amino Acid Sequence; D000818:Animals; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D053782:Transforming Growth Factor beta3 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology
D003470:Culture Media; D011994:Recombinant Proteins; C509044:TGFB3 protein, human; D053782:Transforming Growth Factor beta3
Bioreactor process; Chinese hamster ovary (CHO) cells; Recombinant protein expression; Stable cell line; Transforming growth factor beta 3 (TGF β3)