title: Construction and characterization of stably transfected BHK-21 cells with human-type sialylation characteristic.
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.
authors.affiliation: Department of Protein Glycosylation, GBF-Gesellschaft für Biotechnologische Forschung, Mascheroder Weg 1, D-38124, Braunschweig, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Schlenke
authors.forename: E
authors.initials: E
authors.lastname: Grabenhorst
authors.forename: M
authors.initials: M
authors.lastname: Nimtz
authors.forename: H S
authors.initials: HS
authors.lastname: Conradt