Method for producing the subunits of human interleukin 35 from interleukin 12 and interleukin 27

The method according to any one of the preceding claims, wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:

• 3 and 4) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 12 (SEQ ID NO:
• 3) at sequence position 96, preferably wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 12 (SEQ ID NO:
• 3) at sequence position 96 to any other amino acid than cysteine, preferably to serine or alanine, more preferably to serine, preferably wherein the nucleotide sequence further comprises a nucleotide sequence encoding an affinity tag, and/or
• wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the β-subunit of human Interleukin 12 (SEQ ID NO:
• 4) at sequence position 199, preferably wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the β-subunit of human Interleukin 12 (SQ ID NO:
• 4) at sequence position 199 to serine.

The method according to any one of the preceding claims, wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:

• 3 and 4) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 12 (SEQ ID NO:
• 3) at sequence position 96 and introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the β-subunit of human Interleukin 12 (SEQ ID NO:
• 4) at sequence position 199, preferably wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 12 (SEQ ID NO:
• 3) at sequence position 96 to any other amino acid than cysteine, preferably to serine or alanine, more preferably to serine, and introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the β-subunit of human Interleukin 12 (SEQ ID NO:
• 4) at sequence position 199 to any other amino acid than cysteine, preferably to serine, and/or
• wherein step (b) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 27 polypeptide (SEQ ID NOs:
• 5 and 6) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 27 (SEQ ID NO:
• 5) at sequence position 162, preferably wherein step (b) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 27 polypeptide (SEQ ID NOs:
• 5 and 6) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 27 (SEQ ID NO:
• 5) at sequence position 162 to cysteine, preferably wherein the nucleotide sequence further comprises a nucleotide sequence encoding an affinity tag.

The method according to any one of the preceding claims, wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:

• 3 and 4) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 12 (SEQ ID NO:
• 3) at sequence position 96, and wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the β-subunit of human Interleukin 12 (SEQ ID NO:
• 4) at sequence position 199, and wherein step (b) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 27 polypeptide (SEQ ID NOs:
• 5 and 6) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 27 (SEQ ID NO:
• 5) at sequence position 162,
• preferably wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 12 (SEQ ID NO:
• 3) at sequence position 96 to any other amino acid than cysteine, preferably to serine or alanine, more preferably to serine, and wherein step (a) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 12 polypeptide (SEQ ID NOs:
• 3 and 4) a nucleotide sequence mutating the amino acid residue of the β-subunit of human Interleukin 12 (SEQ ID NO:
• 4) at sequence position 199 to any other amino acid than cysteine, preferably to serine or alanine, more preferably to serine, and wherein step (b) comprises introducing into a nucleic acid molecule encoding the recombinant α- and β-subunits of the human Interleukin 27 polypeptide (SEQ ID NOs:
• 5 and 6) a nucleotide sequence mutating the amino acid residue of the α-subunit of human Interleukin 27 (SEQ ID NO:
• 5) at sequence position 162 to cysteine.