Methods and Vectors for Generating Asialylated Immunoglobulins

Abstract

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.

What is to be claimed: 
     
         1 . An expression vector comprising a nucleotide sequence encoding a sialidase enzyme, wherein the vector is capable of expressing a polypeptide having sialidase enzymatic activity in a mammalian cell culture expressing an Fc-containing molecule. 
     
     
         2 . The vector of  claim 1 , wherein the sialidase enzyme is  Arthrobacter ureafaciens  sialidase enzyme A and 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 vector of  claim 1 , wherein the vector comprises a signal sequence, a promoter sequence, and an antibiotic resistance. 
     
     
         4 . The vector of  claim 3 , wherein the signal sequence is a human growth hormone signal sequence, the promoter sequence is a CMV promoter sequence, and the antibiotic resistance is neomycin.