| 主权项 |
1. An in vitro method of altering fucosylation of a human blood mononuclear cell surface, wherein sialyl-Lewis x structures are produced on protein linked O-glycans and N-glycans on said blood mononuclear cell surface and wherein a sialyl-structure of said sialyl Lewis x structures is α3-linked Neu5Ac (N-acetylneuraminic acid), comprising the steps of:
a) culturing, and optionally subculturing at least once, blood mononuclear cells in non-NeuGc (N-glycolylneuranminic acid) and serum free culture medium so that the said sialyl Lewis x structures on said blood mononuclear cell surface protein linked O-glycans consist essentially of only α3-sialylated Lewis x structures, wherein said α3-sialylated Lewis x structures are α3-sialylated N-acetyllactosamines; b) α3-fucosylating said blood mononuclear cells of step a) with guanosine 5′-diphospho-J-L-fucose (GDP-Fuc) and recombinant α1,3-fucosyltransferase enzyme, wherein α3-fucosylation results in formation of α3-fucosylated sialyl-Lewis x structures on said blood mononuclear cell surface, wherein said α3-fucosylated sialyl-Lewis x structures are α3-fucosylated N-acetyllactosamines; c) assaying levels of α3-linked Neu5Ac structures, and optionally α6-linked sialic acid structures, on said blood mononuclear cell surface prior to and/or after step b); and d) contacting said α3-sialylated and α3-fucosylated sialyl-Lewis x structures on said blood mononuclear cell surface with selectin for selectin directed targeting of said blood mononuclear cells. |
