Use of a gene coding for the C4BP protein beta chain for producing recombinant dimeric proteins
AssigneeUniversite De Reims Champagne-Ardenne
InventorsJacques Cohen, Stephane Oudin, Xavier Dervillez, Annelise Gimenez, Beatrice Donvito, Marcelle Tonye-Libyh
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Abstract
A method for preparing recombinant dimeric proteins using a nucleic acid coding for the C-terminal fragment of the C4BP protein β chain including amino acid residues 193 to 252 or a functional variant of the fragment, wherein the fragment enables covalent bonding of the heterologous polypeptides to which it is fused. Dimeric proteins obtained by means of the method and cells for carrying out the method are also described.
Claims31
A method for producing a recombinant dimer protein comprising:
- a) Transfecting host cells with a vector allowing expression of a nucleotide sequence coding for a fusion polypeptide, said fusion polypeptide comprising at least the fragment constituted by amino acids in positions 193 to 252 of the β chain of the human C4BP protein or a functional variant of said fragment by deletion, addition or substitution of one or more amino acids, conserving the capacity to form an at least dimeric protein, said fusion polypeptide further comprising a polypeptide which is heterologous to said β chain;
- b) culturing transfected cells under conditions which are suitable for expressing the nucleotide sequence coding for the fusion polypeptide and the covalent association of two fusion polypeptides in vivo to form a dimeric protein;
- c) recovering the dimeric proteins formed.
A method for producing heterodimers, said method comprising:
- a. transfecting host cells with one or more vectors to allow the expression of one or more nucleotide sequences coding for:
- i. a first fusion polypeptide, said fusion polypeptide comprising the 193 to 252 fragment of the β chain of the human C4BP protein or a functional variant of said fragment and a first polypeptide heterologous to said β chain;
- and
- ii. a second fusion polypeptide, said fusion polypeptide comprising at least the 193 to 252 fragment of the β chain of the human C4BP protein or a functional variant of said fragment, and a second polypeptide heterologous to said β chain the sequence of which is different from that of the first heterologous polypeptide;
- b. culturing transfected cells under conditions appropriate for expressing the nucleotide sequence or sequences coding for the first and second fusion polypeptides and association of two fusion polypeptides in vivo to form a heterodimeric protein;
- c. recovering the heterodimeric proteins formed.
A recombinant dimeric protein, characterized in that it is constituted by two fusion polypeptides, each fusion polypeptide comprising at least a fragment constituted by amino acids in positions 193 to 252 of the β chain of the human C4BP protein or a functional variant of said fragment by deletion, addition or substitution of one or more amino acids conserving the capacity to form an at least dimeric protein, said fusion polypeptide further comprising a polypeptide heterologous to said β chain.
A recombinant heterodimeric protein, characterized in that it is constituted by a first and a second fusion polypeptide, each fusion polypeptide comprising at least one fragment constituted by amino acids in positions 193 to 252 of the β chain of the human C4BP protein or a functional variant of said fragment by deletion, addition or substitution of one or more amino acids conserving the capacity to form a dimeric protein, said first fusion polypeptide further comprising a polypeptide heterologous to said β chain, and the second fusion polypeptide comprising a second polypeptide heterologous to said β chain which differs from the first heterologous polypeptide.
Use, in a method for producing a recombinant dimeric protein, of a nucleic acid coding for a fusion polypeptide, said fusion polypeptide comprising at least the fragment constituted by amino acids in positions 193 to 252 of the β chain of the human C4BP protein or a functional variant of said fragment by addition, deletion or substitution of one or more amino acids, conserving the capacity to form an at least dimeric protein, said fusion polypeptide further comprising a polypeptide heterologous to said β chain.
Use, in a method for producing a recombinant heterodimeric protein, of:
- a) a first nucleic acid coding for a first fusion polypeptide, said first fusion polypeptide comprising at least the fragment constituted by amino acids in positions 193 to 252 of the β chain of the human C4BP protein or a functional variant of the fragment by addition, deletion or substitution of one or more amino acids, conserving the capacity to form a heterodimeric protein, said first fusion polypeptide further comprising a first polypeptide heterologous to said β chain;
- and b) a second nucleic acid coding for a second fusion polypeptide, said second fusion polypeptide comprising at least the fragment constituted by amino acids in positions 193 to 252 of the β chain of the human C4BP protein or a functional variant of the fragment by addition, deletion or substitution of one or more amino acids, conserving the capacity to form a heterodimeric protein, said second fusion polypeptide further comprising a second polypeptide heterologous to said β chain the sequence of which differs from that of the first heterologous polypeptide.
Dependencies are inferred from the claim text.