Overproduction of recombinant human transforming growth factor beta 3 in Chinese hamster ovary cells

Byungseok Choi; Yujin Lee; Jia Pi; Yongsu Jeong; Kwangehee Baek; Jaeseung Yoon

doi:10.1016/j.pep.2015.02.014

Overproduction of recombinant human transforming growth factor beta 3 in Chinese hamster ovary cells

Byungseok Choi, Yujin Lee, Jia Pi, Yongsu Jeong, Kwangehee Baek, Jaeseung Yoon

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

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 133 mg/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 42.11 pg/ml and specific activity of 1.84 × 10⁷ U/mg.

Original language	English
Pages (from-to)	102-106
Number of pages	5
Journal	Protein Expression and Purification
Volume	110
DOIs	https://doi.org/10.1016/j.pep.2015.02.014
Publication status	Published - Jun 2015

Bibliographical note

Publisher Copyright:
© 2015 Elsevier Inc.

Keywords

Bioreactor process
Chinese hamster ovary (CHO) cells
Recombinant protein expression
Stable cell line
Transforming growth factor beta 3 (TGF β3)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.pep.2015.02.014

Cite this

@article{01dae77ff2394f78a0b87d559bc1a4c1,

title = "Overproduction of recombinant human transforming growth factor beta 3 in Chinese hamster ovary cells",

abstract = "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 133 mg/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 42.11 pg/ml and specific activity of 1.84 × 107 U/mg.",

keywords = "Bioreactor process, Chinese hamster ovary (CHO) cells, Recombinant protein expression, Stable cell line, Transforming growth factor beta 3 (TGF β3)",

author = "Byungseok Choi and Yujin Lee and Jia Pi and Yongsu Jeong and Kwangehee Baek and Jaeseung Yoon",

note = "Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

year = "2015",

month = jun,

doi = "10.1016/j.pep.2015.02.014",

language = "English",

volume = "110",

pages = "102--106",

journal = "Protein Expression and Purification",

issn = "1046-5928",

publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Overproduction of recombinant human transforming growth factor beta 3 in Chinese hamster ovary cells

AU - Choi, Byungseok

AU - Lee, Yujin

AU - Pi, Jia

AU - Jeong, Yongsu

AU - Baek, Kwangehee

AU - Yoon, Jaeseung

PY - 2015/6

Y1 - 2015/6

N2 - 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 133 mg/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 42.11 pg/ml and specific activity of 1.84 × 107 U/mg.

AB - 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 133 mg/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 42.11 pg/ml and specific activity of 1.84 × 107 U/mg.

KW - Bioreactor process

KW - Chinese hamster ovary (CHO) cells

KW - Recombinant protein expression

KW - Stable cell line

KW - Transforming growth factor beta 3 (TGF β3)

UR - http://www.scopus.com/inward/record.url?scp=84924240899&partnerID=8YFLogxK

U2 - 10.1016/j.pep.2015.02.014

DO - 10.1016/j.pep.2015.02.014

M3 - Article

C2 - 25727048

AN - SCOPUS:84924240899

SN - 1046-5928

VL - 110

SP - 102

EP - 106

JO - Protein Expression and Purification

JF - Protein Expression and Purification

ER -

Overproduction of recombinant human transforming growth factor beta 3 in Chinese hamster ovary cells

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this