Claims for Patent: 8,338,088

Summary for Patent: 8,338,088

Title:	Methods related to cell surface glycosylation
Abstract:	The present disclosure provides methods for assessing the glycosylation of a target glycoprotein produced by a cell through analysis of cell-surface glycans on the cell. The present disclosure therefore teaches that glycosylation of cell surface proteins can serve as a proxy for glycosylation of other proteins.
Inventor(s):	Collins; Brian Edward (Arlington, MA), Bosques; Carlos J. (Arlington, MA), Zhu; Xiangping (North Grafton, MA), Bulik; Dorota A. (Malden, MA), Thiruneelakantapillai; Lakshmanan (Boston, MA), Parsons; Ian Christopher (Belmont, MA), Shriver; Zachary (Cambridge, MA), Chillakuru; Rajeev (Cambridge, MA), Venkataraman; Ganesh (Bedford, MA)
Assignee:	Momenta Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	12/595,902
Patent Claims:	1. A method of determining one or more characteristics of the glycosylation pattern of a recombinant non-cell surface glycoprotein of interest produced by a cell, the method comprising: a) providing a cell that produces the recombinant non-cell surface glycoprotein of interest, which cell has at least one cell-surface glycan attached to the surface of the cell, so that the cell has a cell surface glycosylation pattern; b) liberating at least one cell surface glycan from the cell; c) determining at least one characteristic of the cell surface glycosylation pattern selected from the group consisting of degree of glycosylation site occupancy, identity of at least one linked glycan, relative amounts of linked glycans, complete or partial composition of linked glycans, and combinations thereof; and d) based on the at least one determined characteristic, establishing one or more characteristics of the glycosylation pattern of the recombinant non-cell surface glycoprotein of interest, wherein the determined characteristic correlates with the one or more characteristics of the glycosylation pattern of the non-cell surface glycoprotein of interest. 2. The method of claim 1, wherein the at least one characteristic of the glycosylation pattern comprises a change in glycosylation pattern of the recombinant non-cell surface glycoprotein of interest. 3. The method of claim 1, wherein the step of establishing one or more characteristics comprises establishing an extent of sialylation present in the glycosylation pattern of the recombinant non-cell surface glycoprotein of interest. 4. The method of clam 1, wherein the step of liberating comprises exposing the cell to a protease. 5. The method of claim 4, wherein the step of exposing comprises subjecting the cell to protease treatment such that at least one cell surface glycoprotein is liberated, wherein the protease treatment does not substantially rupture cell membranes. 6. The method of claim 1, wherein the step of liberating comprises: a) cleaving one or more protein-linked cell-surface glycans from cell surface glycoproteins; and b) characterizing the one or more cleaved protein-linked cell-surface glycans. 7. The method of claim 6, wherein the protein-linked cell-surface glycan is an N-linked glycan. 8. The method of claim 6, wherein the protein-linked cell-surface glycan is an O-linked glycan. 9. The method of claim 6, wherein the protein-linked cell-surface qlycans comprise at least one sialic acid residue. 10. The method of claim 1, further comprising a step of recording information about at least one of the glycosylation patterns in a fixed medium. 11. The method of claim 1, wherein the recombinant non-cell surface glycoprotein of interest is selected from the group consisting of human somatropin, coagulation factor VIIa, coagulation IX, interferon alphacon-1, insulin glargine, and insulin. 12. The method of claim 1, wherein the recombinant non-cell surface glycoprotein of interest is a recombinant antibody. 13. The method of claim 12, wherein the recombinant antibody is selected from the group consisting of etanercept, infliximab, adalimumab, basiliximab, daclizumab, omalizumab, gemtuzumab, alemtuzumab, rituximab, cetuximab, bevacizumab, palivizumab, and abciximab. 14. The method of claim 1, wherein the recombinant non-cell surface glycoprotein of interest is a therapeutic protein product. 15. The method of claim 1, wherein the recombinant non-cell surface glycoprotein of interest is a commercial glycoprotein that is produced industrially. 16. The method of claim 14, wherein the therapeutic protein product is or comprises a product selected from the group consisting of: a hematologic agent, an interferon, a colony stimulating factor, an antibody, an enzyme, and a hormone. 17. The method of claim 16, wherein the therapeutic protein product is or comprises an antibody. 18. The method of claim 14, wherein the therapeutic protein of interest is characterized in that it has undergone regulatory review in one or more countries. 19. The method of claim 14, wherein the at least one characteristic of the glycosylation pattern of the therapeutic protein of interest is compared to a reference sample that is a pharmaceutical product which has an established glycosylation pattern. 20. The method of claim 19, wherein the surface glycosylation pattern of the cell has at least a 75% correlation to the established glycosylation pattern of the reference sample. 21. The method of claim 1, wherein the step of determining the at least one characteristic of the cell surface glycosylation pattern includes a mass spectrometry technique. 22. The method of claim 1, wherein the glycosylation pattern includes one or more glycan structures selected from the group consisting of high mannose structures, hybrid structures, phosphorylated high mannose, sialylated termini, N-acetylneuraminic acid, N-glycolylneuraminic acid, extension on .alpha.1,6 mannose branches, extension on .alpha.1,3 mannose branches, core fucosylation, sulfated glycans, phosphorylated glycans, sialic acid linked to an N-acetylglucosamine, and acetylated glycans.

Details for Patent 8,338,088

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Eli Lilly And Company	HUMATROPE	somatropin	For Injection	019640	06/23/1987		2027-04-16
Eli Lilly And Company	HUMATROPE	somatropin	For Injection	019640	10/16/1986		2027-04-16
Eli Lilly And Company	HUMATROPE	somatropin	For Injection	019640	02/04/1999		2027-04-16
Emd Serono, Inc.	SAIZEN	somatropin	For Injection	019764	10/08/1996		2027-04-16
Emd Serono, Inc.	SAIZEN	somatropin	For Injection	019764	08/29/2000		2027-04-16
Emd Serono, Inc.	SAIZEN	somatropin	For Injection	019764	01/16/2007		2027-04-16