Anti-Dengue Virus (Clone: DENV-2D22)

Sequenced from human survivors of who had experienced a DENV infection during travel to an endemic region.

This recombinant 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.

Recombinant 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 year. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≥ -70°C. Avoid Repeated Freeze Thaw Cycles.

Standard Overnight on Blue Ice.

ELISA
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DENV-2D22 activity is DENV-2 specific and directed against the E homodimer at the DIII + glycan loop with serotype specificity on one E protein and DII around the fusion loop on the other E protein.

Antibody clone DENV-2D22 was identified as strongly neutralizing, capable of inhibiting infection of DENV-2, and able to bind intact DENV-2 but not DIII or recombinant E^3,4. DENV-2D22 also did not bind to E protein by immunoblot⁴. DENV-2D22 is E protein specific based on an escape mutant of DIII at R323G ³ and recognizes a complex quaternary epitope displayed on the intact virus formed by DIII and DII on two different monomers within a single dimer⁵. When the entire DENV-2 DIII region was inserted into the backbone sequence of a DENV-4 molecular clone to create a recombinant virus, DENV-2D22 was capable of binding and neutralization⁶. Only five contact residues differ between DENV-2 and -4 in DIII, and these are likely critical for binding and neutralization⁵.

DENV-2D22 had no detectable ability to enhance DENV infection at a typical concentration range in a Ab-dependent enhancement assay⁴, was capable of binding and neutralizing chimeric yellow fever-dengue vaccine virus serotype 2², and blocked viral infection of viremic blood for Ae. aegypti⁷.

Dengue virus (DENV) is the most common insect-transmitted virus to target humans, with an estimated 390 million infections annually¹. DENVs are members of the Flaviviridae family and can be divided into four closely related but antigenically distinct serotypes². They encode a single-stranded positive sense RNA genome and display 180 copies of envelope (E) glycoprotein and premembrane/membrane (prM/M) proteins. E glycoprotein is comprised of three structural domains, DI, DII, and DIII, and exists as a homodimer in the pre-fusion state on the mature virus particle. E undergoes multiple conformation changes during maturation and fusion.

1. Smith SA, de Alwis AR, Kose N, et al. mBio. 4(6):e00873-13. 2013.
2. Lecouturier V, Berry C, Saulnier A, et al. Vaccine. 37(32):4601-4609. 2019.
3. de Alwis R, Smith SA, Olivarez NP, et al. Proc Natl Acad Sci U S A. 109(19):7439-44. 2012.
4. Smith SA, Zhou Y, Olivarez NP, et al. J Virol. 86(5):2665-2675. 2012.
5. Fibriansah G, Ibarra KD, Ng TS, et al. Science. 349(6243):88-91. 2015.
6. Gallichotte EN, Widman DG, Yount BL, et al. mBio. 6(5):e01461-15. 2015.
7. Tuan Vu T, Clapham H, Huynh VTT, et al. PLoS Negl Trop Dis. 13(11):e0007142. 2019.