Viraffinity™ - Virus and Viral Component Isolation

Featured Applications

Plasma pools of Dengue fever
Romain Fragnoud, Marie Flamand, Frederic Reynier, Philippe Buchy, Vasna Duong, Alexandre Pachot, Glaucia Paranhos-Baccala and Frederic Bedin . Differential proteomic analysis of virus enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue. BMC Infectious Diseases (2015) 15:518.

In brief, the article’s authors report a method to compare the proteomes of virion-enriched fractions purified from plasma pools of patients with dengue fever or severe dengue. Virions were purified by ultracentrifugation combined with Viraffinity™. Following in-gel hydrolysis, peptides were analyzed by LC-MS. The article states “A technique based on ultracentifugation (Step 1) followed by concentration using… Viraffinity™, was created to obtain a fraction of plasma enriched with DV particles. A positive signal corresponding to monomeric (60 kDa) and dimeric E protein (120 KDa) was observed in the DV-infected samples, and became more intense after the Viraffinity™ step…Densitometry indicated that the protein complexity of the purified samples was reduced by roughly 350-fold compared to the unpurified samples.”. The authors conclude that the Viraffinity™ based technique of virion-enrichment allowed them to identify two host proteins that have prognostic value for classifying patients with acute dengue who are more likely to develop a severe dengue.

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;

Viraffinity™ from Biotech Support Group was used for the purification of hantavirus strain 76–118 proliferated in Vero E6 cells. Researchers analyzed toll-like receptor TLR4 to investigate anti-hantavirus immunity and demonstrated TLR4 to be an important component of antiviral immunity against HTNV infection via the MyD88-independent signaling pathway.

Human Mouth Virus

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

ViraPrep™ Lambda successfully increased the titre and number of A1 virus particles.ViraPrep™ worked by increasing the amount of viral genome and producing a viral genome band without shearing and with an apparent increase in size. In this article, increasing the amount of confluent lysis and for extracting the viral genome.

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. InAbstracts of the 97th General Meeting of the American Society for Microbiology 1997, American Society for Microbiology, Washington, D.C.

Community water supplies are likely to contain enteric viruses which may lead to sporadic cases, or even epidemics, of such diseases as infectious hepatitis or poliomyelitis. Authors used Virraffinity™ for concentration of poliovirus for RT-PCR detection. The basic method involves using a method for concentrating waterborne viruses from large volumes of water to microlitre volumes, and their subsequent detection by the polymerase chain reaction (PCR). The detection step involves the extraction of RNA, synthesis of complementary DNA, amplification by PCR of target DNA with specific primers, and confirmation through nucleic acid hybridization with a radiolabelled oligonucleotide probe. The PCR technique detected the presence of enteroviruses in spiked as well as in contaminated water samples.

Food:Polio virus type 1 and Calcivirus

Rapid Detection of Foodborne Viruses from Minimally Processed Foods. Microbial Detection Methods. 2000;

Scientists used Viraffinity™ for concentration and purification of viruses from foods like seafood which undergo minimal supervision. In the study, Viraffinity™ bound >90% of polio virus type 1 and calcivirus inoculated into lettuce homogenates using a simple method involving a microspin column before amplification by RT-PCR.

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

Viraffinity™ was used for purification of viruses from potatoes infested by larvae of T. solanivora. Moreover authors also consider using Viraffinity™ to avoid damage to virus particles during gradient centrifugation because it is a water insoluble elastomeric polyelectrolyte. The simple method included mixing a sample of Viraffinity™ with a filtered homogenate AnchV-infected larvae, incubation at room temperature, and centrifugation. The supernatant was discarded, pellet rinsed and centrifuged again. Repeating this step several times, the virus was separated from Viraffinity™ beads by adding PBS.

Patent

Composition and utility patents for Viraffinity™ and related technologies.

A method of concentrating and recovering an enzyme activity from enveloped viruses present in a biological sample, is described. The method comprises contacting the biological sample in a first buffer solution with a virus-binding matrix, such as an anion exchanger matrix, to attach virus particles present in the sample to the matrix, washing the matrix carrying the virus particles with a second buffer solution to remove components interfering with viral enzyme activity, lysing the immobilized virus particles in a third buffer solution and recovering the concentrated viral enzyme activity from the third buffer solution.

Bedin, Frédéric, and Romain Fragnoud. "Method and kit for determining the probability that a patient will develop a severe case of dengue." U.S. Patent Application No. 15/912,730.

In brief, the patent application describes claims determining whether a patient will develop severe dengue based on a blood sample. In the example, virions were purified from plasma pools of patients with dengue fever or severe dengue. For purification, virions were first enriched by ultracentrifugation and then subsequently purified by Viraffinity™. The patent application states “…resuspension is then purified using an insoluble polyelectrolyte, Viraffinity (BioSupportGroup, USA). For this purpose, 200 microliters of an MN buffer (60 mM MES pH 6.5, 150 mM NaCl) are added to the viral suspension along with 100 microliters of Viraffinity. The mixture is incubated for 5 min at room temperature then centrifuged for 10 min at 1000×g, following supplier instructions. The supernatant is removed and the polymer pellet is rinsed 3 times with 200 microliters of MN buffer. …The presence of the virus in the final samples was verified by immunoblotting with a monoclonal antibody directed against the envelope protein of the dengue virus (E protein).”.

“This patent application follows previously reported research whereby Viraffinity™ complements the gold-standard of enrichment using gradient ultracentrifugation. I am very pleased that the exquisite selectivity profile of Viraffinity™ was in-part the reason the patent’s authors were able to obtain an excellent viral protein signal by Western blot” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.

Performance:

Ratio refers to the volumetric ratio of Viraffinity™ to sample.

^a Tissue culture supernatants containing 1-10% Fetal Bovine Serum.

^b Based on infectivity.

MSDS:
MSDS-Viraffinity™, Catalog Order # V0162