Related Experiment Video
Updated: Jun 28, 2026
High-Throughput Expression and Purification of Human Solute Carriers for Structural and Biochemical Studies
Published on: September 29, 2023
Construction and characterization of stably transfected BHK-21 cells with human-type sialylation characteristic
P Schlenke1, E Grabenhorst, M Nimtz
1Department of Protein Glycosylation, GBF-Gesellschaft für Biotechnologische Forschung, Mascheroder Weg 1, D-38124, Braunschweig, Germany.
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.
