N-acetylhexosamine-containing N-glycans in glycoprotein products

Patent metadata

Field	Value
Publication number	US-9890410-B2
Application number	US-201514848768-A
Country	US
Kind code	B2
Filing date	Sep 9, 2015
Priority date	Mar 12, 2011
Publication date	Feb 13, 2018
Grant date	Feb 13, 2018

Abstract

The present invention provides methods of evaluating a glycoprotein preparation for the absence, presence or amount of an N-acetylhexosamine glycan, e.g., an N-acetylglucosamine glycan.

First claim

The invention claimed is:

1. A method for manufacturing an antibody preparation, the method comprising:
culturing mammalian cells to produce a recombinant antibody preparation;
determining a level of an N-linked glycan consisting of a single N-acetylglucosamine residue in the produced preparation; and
processing the antibody preparation as a pharmaceutical product based upon the determination.

2. The method of claim 1 , further comprising comparing the determined level of N-linked glycans consisting of a single N-acetylglucosamine residue to a reference value for the antibody preparation for the level of N-linked glycans consisting of a single N-acetylglucosamine residue.

3. The method of claim 1 , wherein the determining step comprises the use of a method for identifying or quantifying N-linked glycans consisting of a single N-acetylglucosamine residue selected from the group consisting of: chromatographic methods, mass spectrometry (MS) methods, electrophoretic methods, nuclear magnetic resonance (NMR) methods, monosaccharide analysis, fluorescence methods, UV-VIS absorbance, enzymatic methods, use of a detection molecule, and combinations thereof.

4. The method of claim 1 , wherein the cells are cultured in a bioreactor.

5. The method of claim 1 , wherein the processing step comprises disposing the antibody preparation into a container and the container is a gas or liquid tight container.

6. The method of claim 1 , wherein the cells are CHO cells.

7. The method of claim 1 , wherein the processing step comprises one or more of: formulating the antibody preparation; processing the antibody preparation into a drug product; combining the antibody preparation with a second component; lyophilizing the antibody preparation.

8. The method of claim 1 , wherein the processing step comprises formulating the antibody preparation.

9. The method of claim 1 , wherein the processing step comprises processing the antibody preparation into a drug product.

10. The method of claim 1 , wherein the processing step comprises lyophilizing the antibody preparation.

11. The method of claim 1 , wherein the processing step comprises one or more of: formulating the antibody preparation; processing the antibody preparation into a drug product; combining the antibody preparation with a second component; changing the concentration of the antibody in the preparation; lyophilizing the antibody preparation; combining a first and second aliquot of the antibody to provide a third, larger, aliquot; dividing the antibody preparation into smaller aliquots; disposing the antibody preparation into a container; packaging the antibody preparation; associating a container comprising the antibody preparation with a label; shipping or moving the antibody preparation to a different location.

12. The method of claim 1 , wherein the antibody is a therapeutic antibody.

13. The method of claim 1 , wherein the antibody preparation comprises an N-linked glycan consisting of a single N-acetylglucosamine residue.

14. The method of claim 1 , wherein the antibody preparation comprises an N-linked glycan consisting of a single N-acetylglucosamine residue linked to the Fc domain of the antibody.

15. The method of claim 1 , wherein the antibody preparation comprises an N-linked glycan consisting of a single N-acetylglucosamine residue linked to Asn297 of the Fc domain of the antibody.

16. A method for manufacturing a fusion protein preparation, the method comprising:
culturing mammalian cells to produce a recombinant fusion protein preparation;
determining a level of an N-linked glycan consisting of a single N-acetylglucosamine residue in the produced preparation; and
processing the fusion protein preparation as a pharmaceutical product based upon the determination.

17. The method of claim 16 , further comprising comparing the determined level of N-linked glycans consisting of a single N-acetylglucosamine residue to a reference value for the fusion protein preparation for the level of N-linked glycans consisting of a single N-acetylglucosamine residue.

18. The method of claim 16 , wherein the determining step comprises the use of a method for identifying or quantifying N-linked glycans consisting of a single N-acetylglucosamine residue selected from the group consisting of: chromatographic methods, mass spectrometry (MS) methods, electrophoretic methods, nuclear magnetic resonance (NMR) methods, monosaccharide analysis, fluorescence methods, UV-VIS absorbance, enzymatic methods, use of a detection molecule, and combinations thereof.

19. The method of claim 16 , wherein the cells are cultured in a bioreactor.

20. The method of claim 16 , wherein the processing step comprises disposing the fusion protein preparation into a container and the container is a gas or liquid tight container.

21. The method of claim 16 , wherein the cells are CHO cells.

22. The method of claim 16 , wherein the processing step comprises one or more of: formulating the fusion protein preparation; processing the fusion protein preparation into a drug product; combining the fusion protein preparation with a second component; lyophilizing the fusion protein preparation.

23. The method of claim 16 , wherein the processing step comprises formulating the fusion protein preparation.

24. The method of claim 16 , wherein the processing step comprises processing the fusion protein preparation into a drug product.

25. The method of claim 16 , wherein the processing step comprises lyophilizing the fusion protein preparation.

26. The method of claim 16 , wherein the processing step comprises one or more of: formulating the fusion protein preparation; processing the fusion protein preparation into a drug product; combining the fusion protein preparation with a second component; changing the concentration of the fusion protein in the preparation; lyophilizing the fusion protein preparation; combining a first and second aliquot of the fusion protein to provide a third, larger, aliquot; dividing the fusion protein preparation into smaller aliquots; disposing the fusion protein preparation into a container; packaging the fusion protein preparation; associating a container comprising the fusion protein preparation with a label; shipping or moving the fusion protein preparation to a different location.

27. The method of claim 16 , wherein the fusion protein is a therapeutic fusion protein.

28. The method of claim 16 , wherein the fusion protein preparation comprises an N-linked glycan consisting of a single N-acetylglucosamine residue.

29. The method of claim 16 , wherein the fusion protein preparation comprises an N-linked glycan consisting of a single N-acetylglucosamine residue linked to an Fc domain of the fusion protein.

30. The method of claim 16 , wherein the fusion protein preparation comprises an N-linked glycan consisting of a single N-acetylglucosamine residue linked to Asn297 of an Fc domain of the fusion protein.

31. The method of claim 1 , comprising processing the antibody preparation as a pharmaceutical product if the level of N-linked glycan consisting of a single N-acetylglucosamine residue in the produced preparation is at least 0.5%.

32. The method of claim 1 , comprising processing the antibody preparation as a pharmaceutical product if the level of N-linked glycan c

Assignees

• Momenta Pharmaceuticals Inc

Inventors

• Washburn Nathaniel
• Arevalo Enrique
• Millea Kevin
• Bosques Carlos J
• Duffner Jay
• Collins Brian Edward

Classifications

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C12P21/005 Primary

  Glycopeptides, glycoproteins · CPC title

• C07K2317/70

  characterized by effect upon binding to a cell or to an antigen · CPC title

• G01N33/15

  Medicinal preparations {; Physical properties thereof, e.g. dissolubility} · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title