BIOMAT Database Logo BIOMAT Database Logo

Expression, purification, and bioassay of human stanniocalcin from baculovirus-infected insect cells and recombinant CHO cells. 1998

J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Department of Protein Development, Human Genome Sciences, Inc., 9410 Key West Avenue, Rockville, Maryland 20850, USA. junli_zhang@hgsi.com

Stanniocalcin is a calcium- and phosphate-regulating glycoprotein hormone that was first described in fish where it functions in preventing hypercalcemia. Human cDNA clones encoding the homolog of stanniocalcin have been recently isolated. In this study, the full-length cDNA coding for human stanniocalcin (hSTC) was cloned into both baculovirus and CHO expression vectors. Recombinant hSTC was then produced efficiently from both baculovirus-infected insect cells and CHO cells in large-scale bioreactors. Purification protocols were developed and used to purify recombinant hSTC from both sources in four chromatography steps. The hSTCs from both expression systems were secreted as glycosylated proteins and as disulfide-linked homodimers. The results from glycosylation studies indicated that stanniocalcin from both sources contained N-linked oligosaccharides but no O-linked sugars. In an in vivo bioassay based on the inhibition of gill calcium transport in fishes, the baculovirus and CHO-expressed protein showed biological activity which is dose dependent. The inhibitory effects of hSTC produced from both systems were essentially equipotent in fishes, despite the differences in glycosylation. Consequently, the precise role of the carbohydrate moiety in recombinant hSTC remains to be determined.

UI MeSH Term Description Entries
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
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
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D004305 Dose-Response Relationship, Drug The relationship between the dose of an administered drug and the response of the organism to the drug. Dose Response Relationship, Drug,Dose-Response Relationships, Drug,Drug Dose-Response Relationship,Drug Dose-Response Relationships,Relationship, Drug Dose-Response,Relationships, Drug Dose-Response
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D006023 Glycoproteins Conjugated protein-carbohydrate compounds including MUCINS; mucoid, and AMYLOID glycoproteins. C-Glycosylated Proteins,Glycosylated Protein,Glycosylated Proteins,N-Glycosylated Proteins,O-Glycosylated Proteins,Glycoprotein,Neoglycoproteins,Protein, Glycosylated,Proteins, C-Glycosylated,Proteins, Glycosylated,Proteins, N-Glycosylated,Proteins, O-Glycosylated
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
D006728 Hormones Chemical substances having a specific regulatory effect on the activity of a certain organ or organs. The term was originally applied to substances secreted by various ENDOCRINE GLANDS and transported in the bloodstream to the target organs. It is sometimes extended to include those substances that are not produced by the endocrine glands but that have similar effects. Hormone,Hormone Receptor Agonists,Agonists, Hormone Receptor,Receptor Agonists, Hormone
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man

Related Publications

J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Expression, purification and characterization of recombinant mouse PC6B from baculovirus-infected insect cells.
February 2004, Protein and peptide letters,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Expression and purification of rat recombinant aminopeptidase B secreted from baculovirus-infected insect cells.
July 2004, Protein expression and purification,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Recombinant pp60c-src from baculovirus-infected insect cells: purification and characterization.
November 1993, Preparative biochemistry,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Purification and characterization of human salivary-gland prokallikrein from recombinant baculovirus-infected insect cells.
May 1992, European journal of biochemistry,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Expression purification and characterization of recombinant human inducible prostaglandin G/H synthase from baculovirus-infected insect cells.
February 1996, Protein expression and purification,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Recombinant human 92-kDa type IV collagenase/gelatinase from baculovirus-infected insect cells: expression, purification, and characterization.
June 1997, Protein expression and purification,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Purification and characterization of recombinant human soluble guanylate cyclase produced from baculovirus-infected insect cells.
June 2009, Protein expression and purification,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Expression and purification of human stromelysin 1 and 3 from baculovirus-infected insect cells.
July 1998, Protein expression and purification,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Recombinant protein expression in baculovirus-infected insect cells.
January 2014, Methods in enzymology,
J Zhang, and P Alfonso, and N R Thotakura, and J Su, and M Buergin, and D Parmelee, and A W Collins, and M Oelkuct, and S Gaffney, and S Gentz, and D P Radman, and G F Wagner, and R Gentz
Expression and purification of human matrilysin produced in baculovirus-infected insect cells.
May 1996, Journal of biotechnology,
Copied contents to your clipboard!