The present invention relates to methods of facilitating the expression of recombinant polypeptides from cells,
extracellular fluids,
extracellular fibers, or any combination thereof, obtained from transgenic
insect cells and larvae comprising a bacterial GlcNAc-6-P 2′-epimerase (GNPE), which is capable of converting N-acetyl-D-
glucosamine-6-
phosphate (GlcNAc-6-P) to N-acetyl-D-
mannosamine-6-
phosphate (ManNAc-6-P). The invention relates to methods to promote efficient
glycoconjugate sialylation, by providing simpler ways to produce large
intracellular pools of
sialic acid precursors. The invention is also directed to nucleic acids, vectors, and cells comprising nucleic acids encoding polypeptides involved in the synthesis of
sialic acid precursors, and cells in combination with nucleic acids encoding glycosyltransferases, including sialyltransferases, to facilitate the production of humanized recombinant glycoproteins. The engineered cells can be used to produce glycosylated proteins lepidopteran insects and
cultured cell lines derived from
Spodoptera frugiperda, Trichoplusia ni, and silkworms, such as
Bombyx mori, particularly those that can be infected by
baculovirus expression vectors.