Anti-Murine Norovirus Capsid (Clone A6.2) – Purified in vivo GOLD™ Functional Grade

Brain homogenate containing MNV-1

This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.

Liquid

Functional grade preclinical antibodies are manufactured in an animal free facility using only in vitro protein free cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.

Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.

Research Use Only

2 – 8° C Wet Ice

A6.2 activity is directed against the P domain of the mouse norovirus capsid. The A6.2 epitope maps to the AʹBʹ and EʹFʹ loops of the P2 subdomain. A6.2 binds to the human- mouse norovirus consensus peptide sequence GWWEDHGQL, which aligns with residues 327 to 335 of P2.

Norovirus, a Caliciviridae virus made up of a single major capsid protein (VP1), causes acute gastroenteritis during infection¹. The capsid protein is composed of three structural domains: N (N terminus), S (shell), and P (protruding), with the latter further divided into P1 and P2 subdomains. P1 has moderate sequence diversity, while P2 is highly variable. Murine norovirus (MNV-1) is the first norovirus used to study the immune response in animal models and can infect the intestinal tract of mice following oral inoculation². MNV-1 can infect macrophage-like cells in vivo and can be cultured in primary dendritic cells and macrophages.

A6.2 was generated from an MNV-1-seropositive 129 mouse injected with brain homogenate containing MNV-1². The spleen was harvested, hybridoma fusion performed, and supernatants screened by ELISA for binding to recombinant MNV-1 capsid.

A6.2 is a neutralizing antibody used for structural analysis of MNV in cryo-EM^1,3,4,5,6 and NMR⁷ studies. Neutralization by A6.2 has also been demonstrated in plaque based assays². A6.2 Fab can also neutralize MNV, albeit with 100 times lower efficacy than the intact antibody, showing that neutralization does not require bivalent binding. Additionally, neutralization of MNV with A6.2 Fab does not induce major conformational changes in the virion. Binding of glycochenodeoxycholic acid to MNV abrogates the neutralization capacity of A6.2^6,7. A6.2 is thought to neutralize MNV-1 infection by preventing virion attachment to the cell surface³.

Phage-display oligopeptide library screens have been used to map the binding epitope to the P2 subdomain⁸. A6.2 does not react with capsid protein in Western blot analysis, and therefore likely binds to a 3D epitope¹.

Mouse norovirus can be cultured in cells of the innate immune system, including primary dendritic cells and macrophages.

Host receptor CD300LF, bile acids

1 Katpally U, Wobus CE, Dryden K, et al. J Virol. 82(5):2079-2088. 2008.
2 Wobus CE, Karst SM, Thackray LB, et al. PLoS Biol. 2(12):e432. 2004.
3 Taube S, Rubin JR, Katpally U, et al. J Virol. 84(11):5695-5705. 2010.
4 2Kolawole AO, Li M, Xia C, et al. J Virol. 88(8):4543-4557. 2014.
5 3Kolawole AO, Smith HQ, Svoboda SA, et al. mSphere. 2(5):e00334-17. 2017.
6 Williams AN, Sherman MB, Smith HQ, et al. J Virol. 95(13):e0017621. 2021.
7 Creutznacher R, Maass T, Dülfer J, et al. Commun Biol. 5(1):563. 2022.
8 Lochridge VP, Hardy ME. J Virol. 81(22):12316-12322. 2007.
9 Kolawole AO, Xia C, Li M, et al. J Gen Virol. 95(Pt 9):1958-1968. 2014.
10 Williams AN, Sherman MB, Smith HQ, et al. J Virol. 95(22):e0147121. 2021.