Expression vector for animal cells including CSP-B 5′-SAR factor and method for producing recombinant proteins using same
The present invention relates to an expression vector for animal cells, comprising: (a) CSP-B (Cytotoxic Serine Protease-B) 5′-SAR (Scaffold or Matrix Attachment Region); (b) a promoter operable in animal cells; and (c) a polyadenylation sequence, and to a method for producing recombinant proteins using same. The vector of the present invention includes CSP-B 5′-SAR, and thus has the effect of overcoming the inhibition of gene expression according to the position of a foreign gene introduced into an animal cell, and significantly improving the expression rate of a target protein. The vector of the present invention effectively expresses recombinant proteins for drugs or antibodies in animal cells. The vector of the present invention and the method for producing recombinant proteins using same may be very usefully applied to the industrial mass production of drugs.
1. An expression vector for animal cells, comprising:
a protein-coding nucleotide sequence to be expressed, wherein the protein is a Novel Erythropoiesis Stimulating Protein (NESP) or an anti-human epidermal growth factor receptor 2 (anti-HER2) antibody;
a 5′-scaffold attachment region (5′-SAR) of cytotoxic serine protease-B (CSP-B), wherein the 5′-SAR is present in two copies downstream of the protein-coding nucleotide sequence to be expressed, and the two copies of the 5′-SAR are consecutively present in the vector;
a promoter operable in animal cells; and
a polyadenylation sequence.
2. The expression vector according to claim 1 , wherein the promoter operable in the animal cells is a cytomegalovirus (CMV) promoter, an adenovirus late promoter, a vaccinia virus 7.5K promoter, a Simian virus 40 (SV40) promoter, an SV40E1 promoter, a herpes simplex virus (HSV) tk promoter, a respiratory syncytial virus (RSV) promoter, an elongation factor 1-α (EF1-α) promoter, a metallothionein promoter, a β-actin promoter, a human interleukin-2 (IL-2) gene promoter, a human interferon (IFN) gene promoter, a human IL-4 gene promoter, a human lymphotoxin gene promoter, or a human granulocyte-macrophage colony-stimulating factor (GM-CSF) gene promoter.
3. The expression vector according to claim 1 , wherein the polyadenylation sequence is a bovine growth hormone polyadenylation sequence, an HSV thymidine kinase (TK) polyadenylation sequence, or an SV40 polyadenylation sequence.
4. The expression vector according to claim 1 , wherein the animal cells are Chinese hamster ovary (CHO) cells, VERO cells, HeLa cells, WI38 cells, baby hamster kidney (BHK) cells, COS cells, or Madin-Darby canine kidney (MDCK) cells.
5. An isolated animal cell transfected with the expression vector according to claim 1 .
6. A method for preparing a recombinant protein, comprising incubating the transfected animal cell of claim 5 and expressing the NESP or anti-HER2 antibody.