Cells, vectors and methods for producing biologically active heterodimeric proteins

Abstract

Biologically active heteropolymeric protein composed of a plurality of subunits, both subunits being synthesized in a single cell having an expression vector comprising heterologous DNA encoding the subunits. Preferably the protein is similar to the human or ungulate fertility hormones, LH and FSH.

What is claimed is:  
     
         1 . An expression vector encoding two subunits of a heterodimeric protein, each under the control of a separate promoter, said protein being selected from the group consisting of) luteinizing hormone (LH), follicle stimulating hormone (FSH), and chorionic gonadotropin (CG).  
     
     
         2 . The expression vector of  claim 1 , said expression vector comprising a plasmid.  
     
     
         3 . The expression vector of  claim 1 , said expression vector comprising a replicating virus.  
     
     
         4 . A mammalian cell consisting essentially of a cell transfected by a first expression vector, said transfected cell being capable of producing a biologically active heterodimeric protein having first and second subunits, said selected protein being selected from the group consisting of luteinizing hormone (LH), follicle stimulating hormone (FSH), and chorionic gonadotropin (CG), each of said subunits of said protein being encoded by said first expression vector under the control of a separate promoter, or progeny of said transfected cell containing the promoters and DNA sequences for said subunits imparted by said vector.  
     
     
         5 . The cell of  claim 4  wherein, in said transfected cell, said expression vector is autonomously replicating.  
     
     
         6 . A mammalian cell consisting essentially of a cell transfected by a first expression vector and a distinct second expression vector, said transfected cell being capable of producing a biologically active heterodimeric protein having first and second subunits, said protein being selected from the group consisting of luteinizing hormone (LH), follicle stimulating hormone (FSH), chorionic gonadotropin (CG) and thyroid stimulating hormone (TSH), the first subunit of said protein being encoded by said first expression vector and the second subunit of said protein being encoded by said second expression vector, or progeny of said transfected cell containing the promoters and DNA sequences for said subunits imparted by said vectors.  
     
     
         7 . The cell of  claim 6  wherein, in said transfected cell, said expression vectors are autonomously replicating.  
     
     
         8 . The cell of  claim 4  or  claim 6 , wherein, in said transfected cell, said first vector is a plasmid.  
     
     
         9 . The cell of  claim 4  or  claim 6 , wherein, in said transfected cell, transcription of each of the two different subunits of said heterodimer is under the control of an SV40 late promoter.  
     
     
         10 . The cell of  claim 4  or  claim 6 , wherein, in said transfected cell, transcription of one subunit of said heterodimer is under the control of the SV40 early promoter, and transcription of the other subunit of said heterodimer is under the control of the mouse metallothionein promoter.  
     
     
         11 . The cell of  claim 10 , wherein, in said transfected cell, said first vector comprises at least the 69% transforming region of the bovine papilloma virus genome.  
     
     
         12 . The cell of  claim 4  or  claim 6 , wherein said transfected cell is a monkey cell.  
     
     
         13 . The cell of  claim 4  or  claim 6 , wherein said transfected cell is a mouse cell.  
     
     
         14 . A mammalian cell in accordance with either one of  claim 4  or  claim 6 , wherein said first and second subunits are from the same mammalian species.  
     
     
         15 . A method for producing a biologically active heterodimeric protein comprising culturing the mammalian cells of  claim 4  or  claim 6 .  
     
     
         16 . The cell of  claim 4  or  claim 6 , said protein being luteinizing hormone.  
     
     
         17 . The cell of  claim 4  or  claim 6 , said protein being follicle stimulating hormone.  
     
     
         18 . The cell of  claim 4  or  claim 6 , said protein being chorionic gonadotrophin hormone.  
     
     
         19 . A method for producing a biologically active heterodimeric protein comprising culturing the mammalian cells of  claim 5  or  claim 7 .  
     
     
         20 . A mammalian cell consisting essentially of a cell produced by recombinant DNA techniques, said recombinant cell being capable of producing a biologically active hormone selected from luteinizing hormone (LH), follicle stimulating hormone (FSH) and chorionic gonadotrophin (CG), or progeny of said recombinant cell capable of producing said hormone.