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

Protein Expression and Purification
J ZhangR Gentz

Abstract

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 ca...Continue Reading

References

Dec 1, 1992·Molecular and Cellular Endocrinology·G F WagnerH G Friesen
May 1, 1990·Journal of Cellular Biochemistry·D R ThomsenA P Elhammer
Nov 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·P L FelgnerM Danielsen
Aug 11, 1995·Molecular and Cellular Endocrinology·A C ChangR R Reddel
Apr 1, 1995·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·C MesaM Fountoulakis
Mar 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·G F WagnerD H Copp
Jan 1, 1994·Methods in Enzymology·M R Hardy, R R Townsend
Mar 5, 1996·Proceedings of the National Academy of Sciences of the United States of America·H S OlsenG F Wagner
Feb 1, 1997·Protein Expression and Purification·T A ColemanR Gentz
Apr 4, 1997·Molecular and Cellular Endocrinology·G F WagnerC Chiarot
Aug 1, 1996·Nature Biotechnology·N JenkinsD C James

❮ Previous
Next ❯

Citations

Mar 1, 2008·Arteriosclerosis, Thrombosis, and Vascular Biology·Changyi ChenQizhi Yao
Aug 29, 2009·BMC Structural Biology·Daniel M TrindadeJörg Kobarg
Jul 16, 2009·Journal of Leukocyte Biology·Yanlin WangDavid Sheikh-Hamad
Apr 5, 2012·ISRN Endocrinology·Timothy D J RichardsGraham F Wagner
Mar 19, 2002·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Yuji YoshikoNorihiko Maeda
Aug 12, 2006·Journal of Experimental Zoology. Part A, Comparative Experimental Biology·Graham F Wagner, Gabriel E Dimattia
Mar 3, 2005·American Journal of Physiology. Endocrinology and Metabolism·Mark PacigaGraham F Wagner
Oct 30, 1998·Endocrinology·R VargheseG E DiMattia
Oct 13, 2006·American Journal of Physiology. Renal Physiology·Arup ChakrabortyDavid Sheikh-Hamad
Jan 18, 2008·American Journal of Physiology. Renal Physiology·Olga SazonovaGraham F Wagner
Mar 23, 2006·The Journal of Applied Psychology·Hao Zhao, Scott E Seibert
Mar 31, 1999·Endocrinology·Y YoshikoN Maeda
Jan 9, 2001·Developmental Dynamics : an Official Publication of the American Association of Anatomists·S E Stasko, G F Wagner
Sep 12, 2001·American Journal of Physiology. Endocrinology and Metabolism·R GuoL D Quarles
Oct 23, 2003·American Journal of Physiology. Renal Physiology·John KanellisDavid Sheikh-Hamad
Mar 29, 2003·American Journal of Physiology. Heart and Circulatory Physiology·David Sheikh-HamadKeith Youker
Jan 5, 2006·Canadian Journal of Physiology and Pharmacology·Mark PacigaGraham F Wagner
Sep 16, 2004·American Journal of Physiology. Endocrinology and Metabolism·Anthony D GagliardiGabriel E DiMattia
Oct 13, 2004·Peptides·Cherry TanegaGraham F Wagner
Aug 26, 2021·Human Reproduction Update·Alexa BishopGuy S Whitley

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Proceedings of the National Academy of Sciences of the United States of America
H S OlsenG F Wagner
Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research
G F WagnerJ L Renfro
American Journal of Physiology. Endocrinology and Metabolism
Mark PacigaG F Wagner
© 2022 Meta ULC. All rights reserved