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Biotech Support Group Release: Hantaan Virus Purification Using Viraffinity™ to Analyze Innate Immune Response Signaling Pathways


9/14/2012 9:53:33 AM

PRLog (Press Release) - Sep 11, 2012 - MONMOUTH JUNCTION, NJ.– Authors Yu et al published an article titled, “Hantaan Virus Triggers TLR4-Dependent Innate Immune Responses” in the journal, Viral Immunology, which investigated the cause of endothelial cell dysfunction and regulation of toll like receptor viral responses during the innate immune response caused by Hantaan virus. Viraffinity™ from Biotech Support Group was used for the purification of hantavirus strain 76-118 proliferated in Vero E6 cells. In this paper, scientists studied the signaling pathway of TLR4 in HTNV-infected EVC-304 cells. TLR4 mRNA-silenced EVC-304 (EVC-304 TLR4-) cells and EVC-304 cells were supported in RPMI 1640 media with fetal bovine serum, penicillin, streptomycin. Biotech Support Group’s Viraffinity™ was used to purify HTNV strains 7 & 118 in polymyxin B treated Vero E6 cells. HTNV-infected EVC-304 cells had increased amount of adaptor protein TRIF than EVC-304 TLR4-cells. MyD88 expression did not alter. The transcription factors for NF-?B and IRF-3 were translocated from the cytoplasm into the nucleus in HTNV-infected EVC-304 cells, but not in HTNV-infected EVC-304 TLR4- cells. Thus these results suggested that TLR4 may play an important role in the antiviral immunity of the host against HTNV infection through an MyD88-independent signaling pathway.

Viraffinity™ is directly added to the sample that is then mixed and centrifuged. The centrifuged pellet contains polyelectrolyte-bound viruses that can then be recovered using a moderately alkaline pH solution. Viraffinity™ is supplied as a suspension reagent ready for use. Simply pipette the suspension into the sample at the appropriate ratio, typically 1 volume of Viraffinity™; to 4 volumes sample. Viraffinity™ isolates HIV-1, HIV-1, human serum, chimeric human rhinovirus, adenovirus, reovirus type 3, encephalomyocarditis, porcine parvovirus, unclassified entero-picornavirus, coxsackievirus A24, bacteriophage lambda. Viraffinity™ is also supplied as the enabling component of the ViraPrep™ application kits: ViraPrep™ Lambda and ViraPrep™ Mammal.

Characteristics of Viraffinity™

Purifies whole infectious non-enveloped virus & non-infectious enveloped virus

Isolates antigenic virions, enveloped and non-enveloped

Enriches for viral nucleic acids

Prepares viral samples for subsequent detection and analysis

For more information on of Viraffinity™ visit:

http://biotechsupportgroup.com/node/81/

About Biotech Support Group LLC

Biotech Support Group LLC is a leading provider of genomics and proteomics sample preparation products and enrichment reagent kits as well as integrated biotechnology services for life sciences research, biomarker and drug discovery. Based in New Jersey, it’s principal products include: AlbuVoid™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, NuGel™ for passivated silica-based affinity chromatography, and ProCipitate™ & ProPrep™ for nucleic acid isolation. Currently, Biotech Support Group LLC and ProFACT Proteomics Inc., are collaborating on the development of a proteomics platform used in functional profiling for proteomic analysis and a separations method for generating sub-proteomes used in biomarker and functional proteomic prospecting. For more information, go to: www.biotechsupportgroup.com

CONTACT:

Dr.Swapan Roy

Biotech Support Group

1 Deer Park Drive, Suite M,

Monmouth Junction, NJ 08852, USA

732-274-2866

sales@biotechsupportgroup.com

Viraffinity™ References

Polymyxin B treated Vero E6 cells: Hanta Virus

Hai-Tao Yu, Hong Jiang, Ye Zhang, Xue-Ping Nan, Yu Li, Wei Wang, Wei Jiang, Dong-Qiang Yang, Wen-Jing Su, Jiu-Ping Wang, Ping-Zhong Wang, and Xue-Fan Bai.Hantaan Virus Triggers TLR4-Dependent Innate Immune Responses. Viral Immunology.2012;

Corynebcterium diphtheria genome: Human Mouth Virus

Metagenomic Analysis of the Human Mouth Virus Population and Characterization of Two Lytic Viruses, Al-Jarbou, Ahmed, Theses, Dept. of Infection. Immunity and Inflammation.2009

Polio Virus

Ting, W.T. E., E. M. Nielson, and C.C. Tseng. 1997. The use of Viraffinity matrix to concentrate waterborne polioviruses for RT-PCR detection. Abstr. Q-169. p.484. In Abstracts of the 97th General Meeting of the American Society for Microbiology 1997, American Society for Microbiology, Washington, D.C.

Food: Polio virus type 1 and Calcivirus

Rapid Detection of Foodborne Viruses from Minimally Processed Foods. Microbial Detection Methods. 2000; An isometric virus of the potato tuber moth Tecia solanivora (Povolny) (Lepidoptera: Gelechiidae) has a tri-segmented RNA genome Jean-Louis Zeddam et. al Journal of Invertebrate Pathology, Volume 99, Issue 2, October 2008, Pages 204-211

Patent

Composition and utility patents for Viraffinity™ and related technologies.

Suggested References:

Hai-Tao Yu, Hong Jiang, Ye Zhang, Xue-Ping Nan, Yu Li, Wei Wang, Wei Jiang, Dong-Qiang Yang, Wen-Jing Su, Jiu-Ping Wang, Ping-Zhong Wang, and Xue-Fan Bai.Hantaan Virus Triggers TLR4-Dependent Innate Immune Responses. Viral Immunology.2012;

Hepojoki, J. M., T. Strandin, H. Wang, O. Vapalahti, A. Vaheri, and H. Lankinen. The cytoplasmic tails of hantavirus glycoproteins interact with the nucleocapsid protein. Journal of General Virology.2010;91:2341-2350

Guo P, El-Gohary Y, Prasadan K, Shiota C, Xiao X, Wiersch J, Paredes J, Tulachan S, Gittes GK. Rapid and simplified purification of recombinant adeno-associated virus.Journal of Virology Methods. 2012 Aug;183(2):139-46.

Lock M, Alvira M, Vandenberghe LH, Samanta A, Toelen J, Debyser Z, Wilson JM.

Rapid, simple, and versatile manufacturing of recombinant adeno-associated viral vectors at scale.Human Gene Therapy. 2010 Oct;21(10):1259-71.

Dorsett Y, Tuschl T. siRNAs: applications in functional genomics and potential as therapeutics.Nature Reviews Drug Discovery. 2004;3:318–329

Meyer BJ, Schmaljohn CS. Persistent hantavirus infections: characteristics and mechanisms.Trends Microbiology. 2000;8:61–67

Peters CJ, Simpson GL, Levy H. Spectrum of hantavirus infection: hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome. Annual Reviews Medicine. 1999;50:531–545

Zolotukhin S, Byrne BJ, Mason E, Zolotukhin I, Potter M, Chesnut K, Summerford C, Samulski RJ, Muzyczka N. Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Therapy. 1999 Jun;6(6):973-85

Severson, W., L. Partin, C. S. Schmaljohn, and C. B. Jonsson. 1999. Characterization of the Hantaan nucleocapsid protein-ribonucleic acid interaction. Journal of Biological Chemistry. 274:33732-33739

Schmaljohn CS, Dalrymple JM. Analysis of Hantaan virus RNA: evidence for a new genus of bunyaviridae. Virology. 1983;131:482-491


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