Hantaan Virus (HTNV) Purification Using Viraffinity™ To Analyze Innate Immune Response Of Toll Like Receptor Pathways

Hantaan Virus (HTNV) Purification Using Viraffinity™ To Analyze Innate Immune Response Of Toll Like Receptor Pathways

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-κB 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:

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

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
Posted on Date: 
Fri, 09/14/2012