Construction of a Novel CHO Cell Line Coexpressing Human GlycosyItransferases and fusion PSGL-1-Immunoglobulin G

Conclusion

The CHO SSF3 cell line used as host for our DNA transfections has been previously adapted for growth in suspension cultures in serum-free media. The recombinant CHO SSF3 cells efficiently returned to growth in serum-free suspension culture after extended culture in monolayers using serum-supplemented media. This indicated that the preadapted cell phenotype was stable after transfection and selection of heterologous genes. Furthermore, the recombinant PSGL-1-lgG protein expressed by CHO SSF3 previously cotransfected with plasmids encoding both the N-acetyl-D-glucosaminyl-transferase and α1,3-fucosyltransferase presented a “human-like” glycosylation pattern, which was specifically recognized by anti-Lewis^x antibodies. The recombinant PSGL-1 bound to P-selectin, its natural ligand, showing that it was properly glycosylated. The transfection of mammalian cell lines with plamids encoding human glycosyltransferases of defined specificity provides a tool to generate novel stable host cell lines for the production of recombinant proteins with tailored glycosylation pattern.