NS1 Recombinant Proteins Are Efficiently Produced in Pichia pastoris and Have Great Potential for Use in Diagnostic Kits for Dengue Virus Infections

Diagnostics
Mariana F XistoSérgio Oliveira de Paula

Abstract

Dengue is one of the major diseases causing global public health concerns. Despite technological advances in vaccine production against all its serotypes, it is estimated that the dengue virus is responsible for approximately 390 million infections per year. Laboratory diagnosis has been the key point for the correct treatment and prevention of this disease. Currently, the limiting factor in the manufacture of dengue diagnostic kits is the large-scale production of the non-structural 1 (NS1) antigen used in the capture of the antibody present in the infected patients' serum. In this work, we demonstrate the production of the non-structural 1 protein of dengue virus (DENV) serotypes 1-4 (NS1-DENV1, NS1-DENV2, NS1-DENV3, and NS1-DENV4) in the methylotrophic yeast Pichia pastoris KM71H. Secreted recombinant protein was purified by affinity chromatography and characterized by SDS-PAGE and ELISA. The objectives of this study were achieved, and the results showed that P. pastoris is a good heterologous host and worked well in the production of NS1DENV 1-4 recombinant proteins. Easy to grow and quick to obtain, this yeast secreted ready-to-use proteins, with a final yield estimated at 2.8-4.6 milligrams per liter of culture. We reache...Continue Reading

References

Dec 1, 1985·Molecular and Cellular Biology·J M CreggK R Madden
Feb 23, 2000·Analytical Biochemistry·M PopovR A Reithmeier
Mar 9, 2000·Journal of Virology·A A KhromykhE G Westaway
Nov 4, 2000·Clinical and Diagnostic Laboratory Immunology·J GroenA D Osterhaus
Dec 1, 2000·Molecular Biotechnology·J M CreggD R Higgins
Sep 5, 2001·The Journal of Infection·T Solomon, M Mallewa
Dec 31, 2003·Advances in Virus Research·Brett D Lindenbach, Charles M Rice
Jun 7, 2007·Tropical Medicine & International Health : TM & IH·J FarrarUNKNOWN WHO/TDR Dengue Scientific Working Group
Nov 13, 2007·Lancet·Scott B Halstead
Oct 9, 2008·Tropical Medicine & International Health : TM & IH·P PongsumpunM A Dubois
Sep 11, 2009·The Journal of General Virology·Gilda GrardXavier de Lamballerie
May 7, 2011·Nature Reviews. Microbiology·Rosanna W PeelingSutee Yoksan
May 10, 2011·BioTechniques·Marko LõokeArnold Kristjuhan
Jun 7, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Panisadee AvirutnanJohn P Atkinson
Apr 13, 2012·The New England Journal of Medicine·Cameron P SimmonsBridget Wills
Jun 7, 2012·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Mélanie CaronEric Maurice Leroy
Oct 4, 2012·Expert Review of Anti-infective Therapy·Kin Fai Tang, Eng Eong Ooi
Jan 2, 2013·Proceedings of the National Academy of Sciences of the United States of America·Steven T StoddardThomas W Scott
Mar 14, 2013·FEMS Microbiology Reviews·Marizela DelicBrigitte Gasser
Apr 9, 2013·Nature·Samir BhattSimon I Hay
Sep 3, 2013·Protein Expression and Purification·Silvia A CardosoSérgio O De Paula
Oct 4, 2013·BioMed Research International·S Gowri SankarS John Vennison
Nov 5, 2013·Trends in Biotechnology·Oliver SpadiutChristoph Herwig
Dec 3, 2013·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Claudia RabertJorge G Farías
Jan 10, 2014·Microbial Cell Factories·Katrin WeinhandlAndrea Camattari
Jan 16, 2014·Methods in Enzymology·Klaus H Nielsen
Jan 18, 2014·Virus Research·Jaime Henrique AmorimLuís Carlos de Souza Ferreira
Apr 2, 2014·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Daniel WeinackerJorge G Farías
Oct 18, 2014·Clinical Chemistry and Laboratory Medicine : CCLM·Beret E Amundson, Fred S Apple
Nov 13, 2014·Osong Public Health and Research Perspectives·Seok-Min YunWooYoung Choi
Feb 28, 2015·Applied Microbiology and Biotechnology·Verena PuxbaumBrigitte Gasser
Dec 3, 2015·Microbial Cell Factories·Brigitte GasserDiethard Mattanovich

❮ Previous
Next ❯

Methods Mentioned

BETA
glycosylation
enzyme-linked immunosorbent assays
PCR
ELISA
column affinity chromatography
electrophoresis

Software Mentioned

GraphPad
Prism7

Related Concepts

Related Feeds

Bioreactors (ASM)

Bioreactors are important devices or systems that utilize living cells or enzymes for a chemical process. These devices carefully control and monitor the environment factors such as pH, nutrients level, and temperature, which can affect the yield in a given process. Discover the latest research here.