Methods and vectors for generating asialylated immunoglobulins
View Patent ↗The properties of an Fc-containing protein, for example, an antibody, are controlled by altering the sialylation of the oligosaccharides in the Fc region by transfecting the cell line expressing the Fc-containing protein with a vector sequence encoding a sialidase. The modified Fc-containing proteins have therapeutic utility in diseases or conditions in which it is desirable to control the affinity for one or more of the FcγRI, FcγRIIA, and FcγRIIIA receptors, ADCC activity, macrophage or monocyte activation, serum half-life, and avidity.
1. A method for providing a sialidase enzymatic activity in a culture comprising a mammalian cell engineered to express an antibody, comprising transfecting the mammalian engineered cell with a vector encoding a catalytic domain of Arthrobacter ureafaciens sialidase enzyme A, wherein the polypeptide is secreted into the culture along with the antibody, whereby the antibody glycans can be acted upon by the polypeptide sialidase enzymatic activity, and wherein one or more of the avidity for multiply localized target proteins, the macrophage or monocyte activation, and serum half-life of the antibody are altered.
2. The method of claim 1 , wherein the mammalian cell line is selected from the group consisting of COS-1, COS-7, HEK293, BHK21, CHO, BSC-1, Hep G2, 653, SP2/0, 293, HeLa, YB2/0 or Y3, myeloma, or lymphoma cells, or any derivative, immortalized or transformed cell thereof.
3. The method of claim 2 , wherein the antibody is an anti-TNFα antibody and the cell line is C168M.