Protein secretory factor with high secretory efficiency and an expression vector comprising the same

Abstract

The present invention relates to a novel protein secretion factor, a vector including a nucleic acid sequence encoding the protein secretion factor, and a transformed cell into which the vector is introduced. The invention also relates to a method of producing a target protein using the transformed cell including the vector.

1 . A protein secretion factor having an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         2 . The protein secretion factor according to  claim 1 , wherein the protein is an intrinsic protein or a foreign protein. 
     
     
         3 . The protein secretion factor according to  claim 1 , wherein the protein is selected from the group consisting of human growth hormones, serum proteins, antibodies, immunoglobulins, cytokines, α-, β- and γ-interferons, colony-stimulating factors (GM-CSF), platelet-derived growth factors (PDGF), phospholipase-activating protein (PLAP)s, insulins, tumor necrosis factors (TNF), growth factors, hormones, calcitonins, calcitonin gene related peptides (CGRP), enkephalins, somatomedins, erythropoietins, hypothalamic secretion factors, prolactins, chorionic gonadotropins, tissue plasminogen activators, growth hormone releasing peptides (GHRP), thymic humoral factors (THF), asparaginases, arginases, arginine deaminases, adenosine deaminases, peroxide dismutases, endotoxinase, catalases, chymotrypsins, lipases, uricases, adenosine diphosphatases, tyrosinases, bilirubin oxidases, glucose oxidases, glucosidases, galactosidases, glucocerebrosidases, and glucourodinases. 
     
     
         4 . An expression cassette, comprising (i) promoter and (ii) a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, which is linked to a gene encoding a target protein. 
     
     
         5 . A vector for secretory expression of target protein, comprising an expression cassette comprising a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, which is linked to a gene encoding a target protein. 
     
     
         6 . The vector according to  claim 5 , wherein the protein is selected from the group consisting of human growth hormones, serum proteins, antibodies, immunoglobulins, cytokines, α-, β- and γ-interferons, colony-stimulating factors (GM-CSF), platelet-derived growth factors (PDGF), phospholipase-activating protein (PLAP)s, insulins, tumor necrosis factors (TNF), growth factors, hormones, calcitonins, calcitonin gene related peptides (CGRP), enkephalins, somatomedins, erythropoietins, hypothalamic secretion factors, prolactins, chronic chorionic gonadotropins, tissue plasminogen activators, growth hormone releasing peptides (GHRP), thymic humoral factors (THF), asparaginases, arginases, arginine deaminases, adenosine deaminases, peroxide dismutases, endotoxinase, catalases, chymotrypsins, lipases, uricases, adenosine diphosphatases, tyrosinases, bilirubin oxidases, glucose oxidases, glucosidases, galactosidases, glucocerebrosidases, and glucourodinases. 
     
     
         7 . The vector according to  claim 5 , further comprising an expression cassette comprising a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 8, which is linked to a gene encoding a target protein. 
     
     
         8 . The vector according to  claim 5 , wherein the vector is for secretory expression of an antibody. 
     
     
         9 . The vector according to  claim 8 , wherein the vector comprises:
 a) a first expression cassette comprising a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, which is linked to a gene encoding an antibody light chain; and   b) a second expression cassette comprising a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 8, which is linked to an antibody heavy chain.   
     
     
         10 . The vector according to  claim 9 , wherein the protein secretion factor of b) has an amino acid sequence of SEQ ID NO: 3. 
     
     
         11 . The vector according to  claim 9 , wherein the protein secretion factor of a) has an amino acid sequence of SEQ ID NO: 1, and the protein secretion factor of b) has an amino acid sequence of SEQ ID NO: 3. 
     
     
         12 . The vector according to  claim 9 , wherein the antibody light chain has an amino acid sequence of SEQ ID NO: 48, and the antibody heavy chain has an amino acid sequence of SEQ ID NO: 49. 
     
     
         13 . A transformed cell, comprising the vector according to  claim 5 . 
     
     
         14 . The transformed cell according to  claim 13 , wherein the cell is an animal cell. 
     
     
         15 . (canceled) 
     
     
         16 . A method of producing a target protein, comprising:
 i) culturing a transformed cell, into which the vector according to  claim 5  is introduced, to express a target protein and secrete the target protein to the outside of the cell; and   ii) recovering the target protein from a culture or culture supernatant of the cell of step i).   
     
     
         17 . The method according to  claim 16 , further comprising purifying the recovered target protein. 
     
     
         18 . The method according to  claim 16 , wherein the cell is a Chinese Hamster Ovary (CHO) cell. 
     
     
         19 . A method of producing a target protein, comprising:
 i) culturing a transformed cell, into which the vector according to  claim 6  is introduced, to express a target protein and secrete the target protein to the outside of the cell; and   ii) recovering the target protein from a culture or culture supernatant of the cell of step i).   
     
     
         20 . A method of producing a target protein, comprising:
 i) culturing a transformed cell, into which the vector according to  claim 7  is introduced, to express a target protein and secrete the target protein to the outside of the cell; and   ii) recovering the target protein from a culture or culture supernatant of the cell of step i).   
     
     
         21 . A method of producing a target protein, comprising:
 i) culturing a transformed cell, into which the vector according to  claim 9  is introduced, to express a target protein and secrete the target protein to the outside of the cell; and   ii) recovering the target protein from a culture or culture supernatant of the cell of step i).