Author(s): Schlenke P, Grabenhorst E, Nimtz M, Conradt HS
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Abstract The human Golgi enzyme CMP-NeuAc:Gal(beta1-4)GlcNAc-R alpha2,6-sialyltransferase (ST6N) was stably coexpressed with human erythropoietin (EPO) from a BHK-21A cell line. The cell line was characterized with respect to the expression and in vitro activity of the ST6N and the endogenous alpha2,3-sialyltransferase. Detailed structural analysis of the N-linked carbohydrates of the rhuEPO expressed from the new cell line was performed by HPAE-PAD-mapping, MALDI/TOF-MS and methylation analysis after purification of the recombinant protein by immunoaffinity chromatography. This is the first report describing that the human alpha2,6-sialyltransferase is capable of sialylating, apart from Gal(beta1-4)GlcNAc-R, also GalNAc(beta1-4)GlcNAc-R motifs in vivo, which is not the case for the endogenous BHK-cell alpha2,3-sialyltransferase.
This article was published in Cytotechnology
and referenced in Journal of Glycobiology