Optimization of canine interleukin-12 production using a baculovirus insect cell expression system

BMC Research Notes
Cristiane Garboggini Melo de PinheiroGeraldo Gileno de Sá Oliveira

Abstract

Interleukin-12 is an important cytokine in mediating cellular immune responses. Recombinant single-chain canine IL-12 was produced in a baculovirus-insect cell system with the aim of conducting further studies on modulation of immune responses in dogs. To optimize the production of recombinant canine IL-12, a classical baculovirus and a modified vector (chitinase A and v-cathepsin knockout) were used containing a native or an optimized insert of canine IL-12. The optimized IL-12 construct contained the GP64 signal peptide and was synthesized with optimized codons for expression in Trichoplusia ni cells. Dot-blot and Western blot analysis showed the highest production levels of recombinant IL-12 protein by the use of the modified baculovirus vector containing the optimized insert, at a multiplicity of infection of five and at 48 h after infection. The recombinant cytokine was successfully purified and showed a good degree of purity, integrity, folding, and yield, with very little endotoxin contamination. Recombinant canine IL-12 induced IFN-γ in canine lymphocytes, indicating that it was biologically active. Therefore, this study describes an efficient method to produce adequate amounts of biologically active canine IL-12, usefu...Continue Reading

References

May 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·U GublerP C Familletti
Oct 1, 1993·The Journal of Experimental Medicine·M J BrundaM K Gately
Dec 24, 1997·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·F OkanoK Yamada
Jan 7, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·F J MurphyP R Burd
Nov 9, 2000·Veterinary Immunology and Immunopathology·K TakeharaM Nakamura
Nov 17, 2001·In Vitro Cellular & Developmental Biology. Animal·L IkonomouS N Agathos
May 7, 2004·Veterinary Immunology and Immunopathology·Lenita Ramires dos SantosGeraldo Gileno de Sá Oliveira
Oct 19, 2004·Journal of Virological Methods·Stephen A KabaMonique M van Oers
Jan 3, 2006·Veterinary Immunology and Immunopathology·Omar A SaldarriagaPeter C Melby
May 12, 2009·International Journal of Cancer. Journal International Du Cancer·Tien-Fu ChuangRea-Min Chu
Aug 6, 2009·Cell Biology and Toxicology·Richard B HitchmanLinda A King
Nov 7, 2009·Methods in Enzymology·Donald L Jarvis
May 6, 2010·Biotechnology and Applied Biochemistry·Richard B HitchmanLinda A King
Jul 2, 2010·Journal of Bioscience and Bioengineering·Takanori FurutaHideki Yamaji
Aug 10, 2010·Journal of Molecular Biology·Jinquan LuoJacqueline Benson
Aug 4, 2011·Journal of Invertebrate Pathology·Monique M van Oers
Sep 24, 2014·The Journal of General Virology·Monique M van OersJust M Vlak

❮ Previous
Next ❯

Methods Mentioned

BETA
dot-blot
electrophoresis
protein assay
ELISA
PCR
circular dichroism
glycosylation
dot blot

Software Mentioned

GraphPad Prism

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.