Methods for producing subtypes of humanized antibody against interleukin-6 receptor
An antibody subtype (1) which is a subtype of the humanized PM-1 antibody against interleukin-6 receptor (IL-6R) and in which one C-terminal of the heavy chain is Pro-NH 2 (447), and an antibody subtype (2) which is a subtype of the humanized PM-1 antibody against interleukin-6 receptor (IL-6R) and in which both C-terminals of the heavy chain are Pro-NH 2 (447), and a pharmaceutical composition comprising them.
1. A method of producing an antibody subtype 1 comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody, wherein said antibody subtype 1 comprises a heavy chain that has amino acids 1-448 of the amino acid sequence set forth in SEQ ID NO: 1, a heavy chain that has amino acids 1-447 of the amino acid sequence set forth in SEQ ID NO: 1 and a C-terminal that is Pro-NH2, and a light chain that has the amino acid sequence set forth in SEQ ID NO: 2.
2. A method according to claim 1 , wherein said anti-IL-6R antibody is a humanized PM-1 antibody.
3. A method according to claim 1 , wherein said host cell is a mammalian cell.
4. The method of claim 3 , wherein said mammalian cell is a CHO cell.
5. A method according to claim 1 , wherein said host cell is cultured in serum free medium.
6. A method according to claim 1 , wherein said host cell is cultured in medium supplemented with protein hydrolysate.
7. The method of claim 6 , wherein said protein hydrolysate is derived from beef, pork, soy beans, rice, vegetables or fish meat.
8. The method of claim 7 , wherein said protein hydrolysate is a fish meat-derived protein hydrolysate.
9. The method of claim 6 , wherein said protein hydrolysate is a vegetable-derived protein hydrolysate.
10. A method according to claim 1 , further comprising expressing said anti-IL-6R antibody for production of said antibody subtype.
11. The method of claim 10 , further comprising isolating said antibody subtype.
12. The method of claim 11 , further comprising purifying said antibody subtype.
13. The method of claim 10 , further comprising detecting said antibody subtype.
14. A method of producing an antibody subtype comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody, wherein said antibody subtype comprises a heavy chain that has amino acids 1-448 of the amino acid sequence set forth in SEQ ID NO: 1 in which an N-terminal glutamine (Gln) is pyroglutamylated, a heavy chain that has amino acids 1-447 of the amino acid sequence set forth in SEQ ID NO: 1 in which an N-terminal glutamine (Gln) is pyroglutamylated and a C-terminal that is Pro-NH2, and a light chain that has the amino acid sequence set forth in SEQ ID NO: 2.
15. A method of producing an antibody subtype 2 comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody, wherein said antibody subtype 2 comprises two heavy chains that have amino acids 1-447 of the amino acid sequence set forth in SEQ ID NO: 1 and a C-terminal that is Pro-NH2, and a light chain that has the amino acid sequence set forth in SEQ ID NO: 2.
16. A method of producing an antibody subtype comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody, wherein said antibody subtype comprises two heavy chains that have amino acids 1-447 of the amino acid sequence set forth in SEQ ID NO: 1 in which an N-terminal glutamine (Gln) is pyroglutamylated and a C-terminal that is Pro-NH2, and a light chain that has the amino acid sequence set forth in SEQ ID NO: 2.