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Viraffinity™ - Virus and Viral Component Isolation

Base Price: $635.00

Product Code: V1062

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Description

Safety Data Sheet

Purifies whole infectious non-enveloped virus & non-infectious enveloped virus
Prepares viral samples for subsequent detection and analysis
Alternative or complementary to ultracentrifugation
- “…love this vs sucrose cushions!” Maggie L. Bartlett, BS PhD, Johns Hopkins Bloomberg School of Public Health
- or can complement ultracentrifugation, for viral proteomic enrichment 350X ( Fragnoud et al, 2016 http://www.ncbi.nlm.nih.gov/pubmed/26572220)
Isolates antigenic virions, enveloped and non-enveloped
Enriches for viral nucleic acids; detergent, chaotrope, Trizol® compatible

Viraffinity™ is a unique water-insoluble elastomeric polyelectrolyte that has been engineered for the capture and recovery of viruses. It is supplied as a suspension reagent ready for use. Applications include: purification of whole infectious non-enveloped virus, virions, viral components, and sample preparation for subsequent detection and analysis.

Viraffinity™ is directly added to a pre-conditioned sample, is then mixed and centrifuged using standard micro-centrifuges (not ultra). The centrifuged pellet contains polyelectrolyte-bound viruses that can then be recovered using a moderately alkaline pH solution.

Click Here For Viraffinity™ Product Sheet

Related Viraffinity™ Kits

ViraPrep™ Mammal contains Viraffinity™ and all necessary buffers and protocols for mammalian virus and virion isolation.

https://www.biotechsupportgroup.com/ViraPrep-Mammal-p/vpm.htm

ViraPrep™Lambda is a complete application kit containing Viraffinity™ and all the necessary reagents for obtaining high purity DNA from plate or liquid lysates, suitable for amplification, automated fluorescent sequencing and other common molecular biology techniques.

https://www.biotechsupportgroup.com/ViraPrep-Lambda-p/vlk.htm

Product	size	Item No.
Viraffinity™	15ml	V1062-15

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:

Virus	Titer	Ratio	% Bound b
HIV-1 ^a	7x10 ³ TCID₅₀	1:2	96
HIV-1, human serum	7x10 ³ TCID₅₀	1:2	80
Chimeric Human Rhinovirus ^a	10 ⁶ - 10⁸ pfu/ml	1:3	95
Adenovirus (Ad5d1309) ^a	10 ⁶ - 10⁸ pfu/ml	1:3	90
Reovirus Type 3 ^a	10 ⁶ - 10⁸ pfu/ml	1:4	99
Encephalomyocarditis (EMC) ^a	10 ⁷TCID₅₀	1:4	99
Porcine Parvovirus ^a	10 ⁷TCID₅₀	1:2	90
Unclassified Entero-Picornavirus ^a	10 ⁶TCID₅₀	1:4	90
Coxsackievirus A24 ^a	10 ⁶ - 10⁷ pfu/ml	1:2	70-95
Bacteriophage Lambda	10 ⁹ pfu/ml	1:5	>95

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

References

Cell Culture Supernatant Fluids

Yeh, Jane X., et al. "Treatment of Sindbis Virus-Infected Neurons with Antibody to E2 Alters Synthesis of Complete and nsP1-Expressing Defective Viral RNAs ." Mbio (2022): e02221-22.

Since the COVID-19 virus can be detected in stool samples, it has been proposed that detection of the virus in wastewaters will help to monitor local or regional outbreaks. For this to succeed, simple and robust measurement of viruses from wastewater is necessary. Conventionally, enteric viruses are first concentrated by a variety of filter methods. A secondary enrichment step is often employed, to reduce to a final volume and improve purity prior to final analysis. This secondary step may be provided by the Viraffinity™ reagent, and does not require ultracentrifugation. As described in these two references: 1) Viraffinity™ can dramatically improve the purification of viruses, and 2) can remove RT-PCR inhibitors in the detection of enteric viruses.

1) Romain Fragnoud R., et al. "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. http://www.ncbi.nlm.nih.gov/pubmed/26572220

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™. “…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.”.

2) Leggitt, Paris R., and Lee-ann Jaykus. "Detection methods for human enteric viruses in representative foods." Journal of Food Protection 63.12 (2000): 1738-1744.

“To optimize viral nucleic acid amplification, secondary PEG precipitates… required a prior adsorption step with an equal volume of Viraffinity™ to further remove RT-PCR inhibitors. In this case, viruses …were adsorbed by the addition of an equal volume of Viraffinity™, … These Viraffinity™ precipitates were used directly in subsequent RNA extractions.”

Patents

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.

Parthasarathy, Ranjani V., Hsi-Chou Liu, Wensheng Xia, Assumpta Bennaars-Eiden, Paul N. Holt, William Bedingham, Christopher R. Kokaisel, and Jeffrey C. Pederson. " Methods for nucleic acid amplification ." U.S. Patent Application 12/596,346, filed April 25, 2008.

Parthasarathy, Ranjani, Katya Ericson, and William Bedingham. " Methods for nucleic acid isolation and kits using a microfluidic device and sedimenting reagent ." U.S. Patent Application 10/852,022, filed May 24, 2004.

Bedingham, William, et al. " Variable valve apparatus and methods ." U.S. Patent No. 8,057,758. 15 Nov. 2011.

Parthasarathy, Ranjani, Katya Ericson, and William Bedingham. " Methods for nucleic acid isolation and kits using solid phase material ." U.S. Patent Application No. 10/852,645.

Parthasarathy, Ranjani V., Katya Ericson, and William Bedingham. " Methods for nucleic acid isolation and kits using a microfluidic device and concentration step. " U.S. Patent 7,939,249, issued May 10, 2011.

Bedingham, William, Barry Robole, Ranjani Parthasarathy, and Katya Ericson. "Variable Valve Apparatus and Methods." U.S. Patent Application 11/684,656, filed March 12, 2007.

Bedingham, William, et al. " Variable Valve Apparatus and Methods. " U.S. Patent Application No. 11/684,656.

European Patent Application EP2305809 Methods for nucleic acid isolation and kits using a microfluidic device and concentration step

Journal Citations

Dengue Virus
Fragnoud, Romain, et al. " Differential proteomic analysis of virus-enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue ." BMC infectious diseases 15.1 (2015): 518.

Food Samples

Maurer, John, ed. PCR methods in foods . Springer Science & Business Media, 2006.

D’Souza, D. H. " Update on foodborne viruses: types, concentration and sampling methods. " Advances in Microbial Food Safety 2 (2014): 102.

Jaykus, Lee-Ann. " Detection of human enteric viruses in foods. " Foodborne Diseases Handbook 2 (2000): 137-163.

Sair, A. I., D. H. D'souza, and L. A. Jaykus. " Human enteric viruses as causes of foodborne disease ." Comprehensive reviews in food science and food safety 1.2 (2002): 73-89.

Lee, A. " Rapid Detection of Foodborne Viruses from Minimally Processed Foods , poster Australian Society of Microbiology." (2000).

Grohmann, Gary, and Alvin Lee. " Viruses, food and environment." Foodborne microorganisms of public health significance . Sydney: AIFST (1997): 603-620.

Savers, Screen. " EXHIBIT DESCRIPTIONS. " Microbial & Comparative Genomics 2.3 (1997): 244-249.

Use of Viraffinity and Multiplex RT-PCR for the dectection of Food borne viruses

Food and Water Samples

Grohmann, Gary, and Alvin Lee. " Viruses, food and environment." Foodborne microorganisms of public health significance. Sydney: AIFST (1997): 603-620.

Isometric Virus

Zeddam, Jean-Louis, et al. " An isometric virus of the potato tuber moth Tecia solanivora(Povolny)(Lepidoptera: Gelechiidae) has a tri-segmented RNAgenome ." Journal of invertebrate pathology 99.2 (2008): 204-211.

Hantaan Virus

Yu, Hai-Tao, Hong Jiang, Ye Zhang, Xue-Ping Nan, Yu Li, Wei Wang, Wei Jiang et al. " Hantaan virus triggers TLR4-dependent innate immune responses ."Viral immunology 25, no. 5 (2012): 387-393.

Wang, Ping-Zhong, Chang-Xing Huang, Ye Zhang, Zhi-Dong Li, Hai-Tao Yu, Ying Zhang, Zhan-Sheng Jia et al. " Analysis of the immune response to Hantaan virus nucleocapsid protein C-terminal-specific CD8+ T cells in patients with Hemorrhagic fever with renal syndrome ." Viral immunology 22, no. 4 (2009): 253-260.

Jiang, Hong, Ping-Zhong Wang, Ye Zhang, Zhe Xu, Li Sun, Li-Mei Wang, Chang-Xing Huang et al. " Hantaan virus induces toll-like receptor 4 expression, leading to enhanced production of beta interferon, interleukin-6 and tumor necrosis factor-alpha. " Virology 380, no. 1 (2008): 52-59.

Human Enteric Viruses

Leggitt, Paris R., and Lee-ann Jaykus. " Detection methods for human enteric viruses in representative foods ." Journal of Food Protection® 63.12 (2000): 1738-1744.

Jaykus, Lee-Ann. " Detectionof human enteric viruses in foods. " Foodborne Diseases Handbook 2 (2000): 137-163.

Human Mouth Virus

Al-Jarbou, Ahmed. Metagenomic Analysis of the Human Mouth Virus Population and Characterisation of Two Lytic Viruses . Diss. University of Leicester, 2009.

Lepidoptera: Gelechiidae

Zeddam, Jean-Louis, et al. " An isometric virus of the potato tuber moth Tecia solanivora (Povolny)(Lepidoptera: Gelechiidae) has a tri-segmented RNA genome ." Journal of invertebrate pathology 99.2 (2008): 204-211.

Serum

Moreau, Isabelle, John Levis, Orla Crosbie, Elizabeth Kenny-Walsh, and Liam J. Fanning et al. " Correlation between pre-treatment quasispecies complexity and treatment outcome in chronic HCV genotype 3a ." Virology journal 5, no. 1 (2008): 1.

Abdurahman, Samir. Studies on HIV-1 core assembly . Institutionen för laboratoriemedicin/Department of Laboratory Medicine, 2007.

Human Immunodeficiency Virus Type 1

Abdurahman, Samir, Akos Végvári, Masoud Youssefi, Michael Levi, Stefan Höglund, Elin Andersson, Peter Horal, Bo Svennerholm, Jan Balzarini, Anders Vahlne, et al. " Activity of the small modified amino acid α-hydroxy glycineamide on in vitro and in vivo human immunodeficiency virus type 1 capsid assembly and infectivity ." Antimicrobial agents and chemotherapy 52, no. 10 (2008): 3737-3744.

Abdurahman, S., Höglund, S., Höglund, A., & Vahlne, A et al,. (2007). Mutation in the loop C-terminal to the cyclophilin A binding site of HIV-1 capsid protein disrupts proper virus assembly and infectivity . Retrovirology, 4(1), 1.

Abdurahman, Samir, Masoud Youssefi, Stefan Höglund, and Anders Vahlne, et al. " Characterization of the invariable residue 51 mutations of human immunodeficiency virus type 1 capsid protein on in vitro CA assembly and infectivity ." Retrovirology 4, no. 1 (2007): 1.

Abdurahman, Samir, Stefan Höglund, Laura Goobar-Larsson, and Anders Vahlne . " Selected amino acid substitutions in the C-terminal region of human immunodeficiency virus type 1 capsid protein affect virus assembly and release ." Journal of general virology 85, no. 10 (2004): 2903-2913.

Plasma

Fragnoud, Romain, 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 15, no. 1 (2015): 518.

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.

Bacteriophage Lambda DNA

Hitti, J. "Fast and Convenient Purification of Bacteriophage Lambda DNA with Viraffinity™ Matrix, poster Cold Spring Harbor Conference on Genome Mapping & Sequencing." (1997).