Title:
SOLUBLE EXPRESSION OF BULKY FOLDED ACTIVE PROTEINS
Document Type and Number:
WIPO Patent Application WO/2011/142529
Kind Code:
A2
Abstract:
The present invention relates to expression vectors and methods for enhancing soluble expression and secretion of a heterologous protein, particularly a bulky folded active heterologous protein which has one or more transmembrane-like domains or intramolecular disulfide bonds by linking a leader peptide with acidic or basic pI and high hydrophilicity thereto; by substituting one or more amino acids within N-terminal of the heterologous protein with ones having acidic or neutral pI and high hydrophilicity; or reducing elevating GRNA value of a polynucleotide encoding the leader peptide having basic pI value and high hydrophilicity. The expression vector and the method may be used to produce of heterologous protein and to transduce of therapeutic proteins in a patient by preventing formation of insoluble inclusion body and by enhancing secretional efficiency of the heterologous protein into the periplasm or outside cell.
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Inventors:
LEE SANG JUN (KR)
KIM YOUNG OK (KR)
NAM BO HYE (KR)
KONG HEE JEONG (KR)
KIM KYUNG KIL (KR)
KIM YOUNG OK (KR)
NAM BO HYE (KR)
KONG HEE JEONG (KR)
KIM KYUNG KIL (KR)
Application Number:
PCT/KR2011/001465
Publication Date:
November 17, 2011
Filing Date:
March 03, 2011
Export Citation:
Assignee:
REPUBLIC OF KOREA REPRESENTED BY NAT FISHERIES RES & DEV INST (KR)
LEE SANG JUN (KR)
KIM YOUNG OK (KR)
NAM BO HYE (KR)
KONG HEE JEONG (KR)
KIM KYUNG KIL (KR)
LEE SANG JUN (KR)
KIM YOUNG OK (KR)
NAM BO HYE (KR)
KONG HEE JEONG (KR)
KIM KYUNG KIL (KR)
International Classes:
C12N15/63; C12N15/11
Foreign References:
| KR20090055457A | 2009-06-02 | |||
| KR20070079025A | 2007-08-03 |
Other References:
STEPHEN R. CAMPION ET AL.: 'Amino-Terminal Charge Affects the Periplasmic Accumulation of Recombinant Heregulin/EGF Hybrids Exported Using the Escherichia coli Alkaline Phosphatase Signal Sequence.' PROTEIN EXPRESSION AND PURIFICATION. vol. 10, no. 3, 1997, pages 331 - 339
LEI ZHANG ET AL.: 'Alteration in the IL-2 signal peptide affects secretion of proteins in vitro and in vivo.' THE JOURNAL OF GENE MEDICINE. vol. 7, no. 3, 2005, pages 354 - 365
N. ABDULLAH ET AL.: 'Removal of Poly-Histidine Fusion Tags From Recombinant Proteins Purified by Expanded Bed Adsorption.' BIOTECHNOLOGY AND BIOENGINEERING. vol. 92, no. 4, 2005, pages 501 - 513
LEI ZHANG ET AL.: 'Alteration in the IL-2 signal peptide affects secretion of proteins in vitro and in vivo.' THE JOURNAL OF GENE MEDICINE. vol. 7, no. 3, 2005, pages 354 - 365
N. ABDULLAH ET AL.: 'Removal of Poly-Histidine Fusion Tags From Recombinant Proteins Purified by Expanded Bed Adsorption.' BIOTECHNOLOGY AND BIOENGINEERING. vol. 92, no. 4, 2005, pages 501 - 513
Attorney, Agent or Firm:
HAN, Yun Ho et al. (Seokcheon Building1551-5, Seocho-dong, Seocho-gu, Seoul 137-873, KR)
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Claims: