Characterization of CHO-MIF gene and protein, and use thereof
View Patent ↗The present invention is concerned with the specific and highly sensitive detection of specific CHO-MIF (macrophage migration inhibitory factor from Chinese Ovarian Hamster cell line) complexes in the production of anti-MIF antibodies. The present invention is further concerned with the provision of specific antibodies which can be used for a CHO-MIF detection method. The present invention is also concerned with a CHO MIF knockout cell line and use thereof. The present invention also provides preparations obtained from recombinant production in CHO cell lines which are essentially free of CHO-MIF.
1. A method for the detection of Chinese hamster ovary (CHO)-macrophage migration inhibitory factor (MIF) contaminations in a monoclonal anti-MIF antibody preparation, the method comprising:
i) contacting the anti-MIF antibody preparation with a polyclonal anti-CHO-MIF antibody, affinity purified against CHO-MIF, and
ii) detecting the presence of CHO-MIF.
2. The method of claim 1 wherein the CHO-MIF contaminates a final CHO cell produced monoclonal anti-MIF antibody —preparation or a preparation of antigen-binding fragments thereof.
3. The method of claim 1 wherein the CHO-MIF is endogenous CHO-MIF produced by CHO cells.
4. The method of claim 1 wherein the detection is carried out by a semi-quantitative Western Blot analysis.
5. A method for detecting CHO-MIF contaminations in a preparation during production of monoclonal anti-MIF antibodies or antigen-binding fragments thereof or in the final preparation of monoclonal anti-MIF antibody or antigen-binding fragments thereof, the method comprising:
i) contacting the monoclonal anti-MIF antibodies or antigen-binding fragments thereof containing preparation with a rabbit anti-CHO-MIF antibody, affinity purified against CHO-MIF, and
ii) detecting the presence of CHO-MIF in the preparation.
6. The method of claim 5 wherein the detecting is carried out as a semi-quantitative Western Blot analysis.
7. A method for the production of anti-macrophage migration inhibitory factor (MIF) antibodies or antigen-binding fragments thereof in Chinese hamster ovary (CHO) cells, wherein the method comprises:
i) producing the anti-MIF antibodies or fragments thereof in a cell culture supernatant,
ii) contacting the cell culture supernatant with a polyclonal anti-CHO-MIF antibody, affinity purified against CHO-MIF, and
iii) detecting the presence of CHO-MIF in the cell culture supernatant.
8. A method for producing a recombinant human macrophage migration inhibitory factor (MIF) antibody preparation, the method comprising;
expressing the recombinant human MIF antibody in a Chinese hamster ovary (CHO) cell line,
ii) contacting the preparation with a polyclonal anti-CHO-MIF antibody, affinity purified against CHO-MIF, and
iii) detecting the presence of CHO-MIF.
9. The method of claim 8 wherein the amount of the CHO-MIF detected is below 0.5 ppm.
10. The method of claim 8 , wherein the recombinant human macrophage MIF antibody is selected from the group consisting of:
i. a RAB9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25111 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25113,
ii. a RAB4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25110 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25112,
iii. a RAB0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25114 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25115,
iv. a RAM9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25859 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25860,
v. a RAM4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25861 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25862,
vi. a RAM0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25863 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25864,
vii. a RAB9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:22 and a heavy chain amino acid sequence of SEQ ID NO:26,
viii. a RAB4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:23 and a heavy chain amino acid sequence of SEQ ID NO:27,
ix. a RAB0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:24 and a heavy chain amino acid sequence of SEQ ID NO:28,
x. a RAM9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:33 and a heavy chain amino acid sequence of SEQ ID NO:32,
xi. a RAM4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:35 and a heavy chain amino acid sequence of SEQ ID NO:34, and
xii. a RAM0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:31 and a heavy chain amino acid sequence of SEQ ID NO:30.
11. The method of claim 1 wherein the amount of the CHO-MIF is below 0.5 ppm.
12. The method of claim 11 wherein the amount of the CHO-MIF is below 0.2 ppm.
13. The method of claim 5 wherein the amount of the CHO-MIF is below 0.5 ppm.
14. The method of claim 13 wherein the amount of the CHO-MIF is below 0.2 ppm.
15. The method of claim 7 wherein the amount of the CHO-MIF is below 0.5 ppm.
16. The method of claim 15 wherein the amount of the CHO-MIF is below 0.2 ppm.
17. The method of claim 9 wherein the amount of the CHO-MIF is below 0.2 ppm.
18. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAB9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25111 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25113.
19. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAB4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25110 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25112.
20. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAB0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25114 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25115.
21. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAM9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25859 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25860.
22. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAM4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25861 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25862.
23. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAM0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25863 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25864.
24. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAB9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:22 and a heavy chain amino acid sequence of SEQ ID NO:26.
25. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAB4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:23 and a heavy chain amino acid sequence of SEQ ID NO:27.
26. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAB0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:24 and a heavy chain amino acid sequence of SEQ ID NO:28.
27. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAM9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:33 and a heavy chain amino acid sequence of SEQ ID NO:32.
28. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAM4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:35 and a heavy chain amino acid sequence of SEQ ID NO:34.
29. The method of claim 10 wherein the recombinant human macrophage MIF antibody is a RAM0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:31 and a heavy chain amino acid sequence of SEQ ID NO:30.