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Tissue plasminogen activator coexpressed in Chinese hamster ovary cells with alpha(2,6)-sialyltransferase contains NeuAc alpha(2,6)Gal beta(1,4)Glc-N-AcR linkages. 1995

S L Minch, and P T Kallio, and J E Bailey
Department of Chemical Engineering, California Institute of Technology, Pasadena 91125, USA.

Genetic alteration of the set of oligosaccharide biosynthesis enzymes expressed in a genetically engineered host cell line is a plausible strategy for manipulating the oligosaccharides on a cloned glycoprotein coexpressed in that cell line. This hypothesis was verified for the particular case of sialylation of recombinant human tissue plasminogen activator (tPA) expressed by an engineered Chinese hamster ovary (CHO) cell line. The gene for rat liver beta-galactoside alpha(2,6)-sialyltransferase (2,6-ST) was cloned behind the MMTV promoter in the vector pMSG and transfected into a tPA-expressing CHO cell line. Selected and screened transfectants exhibited significantly greater surface fluorescence than controls in flow cytometric analyses of cells labeled with Sambacus nigra agglutinin (SNA)-biotin and streptavidin-R-phycoerythrin; SNA specifically binds to NeuAc alpha(2,6)Gal beta(1,4)Glc-N-AcR linkages, which are synthesized by 2,6-ST and which are not normally found on CHO cells. SNA blots of partially purified tPA from the culture supernatant demonstrated that tPA synthesized in the 2,6-ST transfectants possessed terminal NeuAc alpha(2,6)Gal beta(1,4)Glc-N-AcR linkages, while tPA from the original recombinant CHO cell line did not. Besides possibly allowing the production of glycoproteins in cell culture with glycosylation more closely resembling that in humans, extensions of this strategy have the potential to tailor the pharmacokinetics, targeting, and antigenic properties of cloned glycoproteins.

UI MeSH Term Description Entries
D008562 Membrane Glycoproteins Glycoproteins found on the membrane or surface of cells. Cell Surface Glycoproteins,Surface Glycoproteins,Cell Surface Glycoprotein,Membrane Glycoprotein,Surface Glycoprotein,Glycoprotein, Cell Surface,Glycoprotein, Membrane,Glycoprotein, Surface,Glycoproteins, Cell Surface,Glycoproteins, Membrane,Glycoproteins, Surface,Surface Glycoprotein, Cell,Surface Glycoproteins, Cell
D008956 Models, Chemical Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment. Chemical Models,Chemical Model,Model, Chemical
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D010959 Tissue Plasminogen Activator A proteolytic enzyme in the serine protease family found in many tissues which converts PLASMINOGEN to FIBRINOLYSIN. It has fibrin-binding activity and is immunologically different from UROKINASE-TYPE PLASMINOGEN ACTIVATOR. The primary sequence, composed of 527 amino acids, is identical in both the naturally occurring and synthetic proteases. Alteplase,Plasminogen Activator, Tissue-Type,T-Plasminogen Activator,Tissue-Type Plasminogen Activator,Actilyse,Activase,Lysatec rt-PA,TTPA,Tisokinase,Tissue Activator D-44,Lysatec rt PA,Lysatec rtPA,Plasminogen Activator, Tissue,Plasminogen Activator, Tissue Type,T Plasminogen Activator,Tissue Activator D 44,Tissue Type Plasminogen Activator
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002236 Carbohydrate Conformation The characteristic 3-dimensional shape of a carbohydrate. Carbohydrate Linkage,Carbohydrate Conformations,Carbohydrate Linkages,Conformation, Carbohydrate,Conformations, Carbohydrate,Linkage, Carbohydrate,Linkages, Carbohydrate
D002240 Carbohydrate Sequence The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS. Carbohydrate Sequences,Sequence, Carbohydrate,Sequences, Carbohydrate
D006224 Cricetinae A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS. Cricetus,Hamsters,Hamster
D000094802 beta-D-Galactoside alpha 2-6-Sialyltransferase Sialyltransferases that catalyze the transfer of N-ACETYLNEURAMINIC ACID from CYTIDINE MONOPHOSPHATE N-ACETYLNEURAMINIC ACID to the 6-OH of the GALACTOSE residue of N-GLYCANS. CMP-Acetylneuraminate Galactoside (alpha 2-6)-Sialyltransferase,CMP-N-Acetylneuraminate-Galactosylglycoprotein Sialyltransferase ST6GAL,CMP-N-Acetylneuraminate-beta-Galactoside alpha-2,6-Sialyltransferase,CMP-N-Acetylneuraminic Acid-Lactose Sialytransferase,CMP-NeuAc-Galactoside (alpha 2-6)-Sialyltransferase,CMP-Sialic Acid-N-Acetyllactosaminide alpha (2-6)-Sialyltransferase,Gal-1-4-GlcNAc alpha(2-6)-Sialyltransferase,alpha 2-6-Sialyltransferase,alpha2,6(N) Sialyltransferase,alpha6-Sialyltransferase,beta-D-Galactoside alpha-2-6-Sialyltransferase,beta-Galactoside alpha2,6-Sialyltransferase,beta-Galactosyl(1-4)N-Acetylglucosaminide alpha(2-6)-Sialyltransferase,Gal-GlcNAc(2-6)-sialyltransferase,SIAT-1,ST6(N),ST6Gal I,ST6Gal-1,2-6-Sialyltransferase, alpha,2-6-Sialyltransferase, beta-D-Galactoside alpha,Acid-Lactose Sialytransferase, CMP-N-Acetylneuraminic,CMP N Acetylneuraminate Galactosylglycoprotein Sialyltransferase ST6GAL,CMP N Acetylneuraminate beta Galactoside alpha 2,6 Sialyltransferase,CMP N Acetylneuraminic Acid Lactose Sialytransferase,SIAT 1,ST6GAL, CMP-N-Acetylneuraminate-Galactosylglycoprotein Sialyltransferase,ST6Gal 1,Sialyltransferase ST6GAL, CMP-N-Acetylneuraminate-Galactosylglycoprotein,Sialytransferase, CMP-N-Acetylneuraminic Acid-Lactose,alpha 2 6 Sialyltransferase,alpha 2-6-Sialyltransferase, beta-D-Galactoside,alpha-2,6-Sialyltransferase, CMP-N-Acetylneuraminate-beta-Galactoside,alpha-2-6-Sialyltransferase, beta-D-Galactoside,alpha2,6-Sialyltransferase, beta-Galactoside,alpha6 Sialyltransferase,beta D Galactoside alpha 2 6 Sialyltransferase,beta Galactoside alpha2,6 Sialyltransferase
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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