Development of broadly neutralizing monoclonal antibodies against western equine encephalitis virus (WEEV) and eastern equine encephalitis virus (EEEV)


  1. Hu, W-G.
  2. Nagata, L.P.
  3. Ennis, J.
  4. Zeitlin, L.
Corporate Authors
Defence Research and Development Canada, Suffield Research Centre, Ralston AB (CAN)
Defence Research and Development Canada – Suffield Research Centre is in collaboration with Mapp Biopharmaceutical, Inc. to develop broadly neutralizing monoclonal antibodies (NAbs) against western and eastern equine encephalitis viruses (WEEV and EEEV). In order to develop these NAbs, female Balb/c mice were immunized with immunogens of WEEV or EEEV in different formats with different strains. After three immunizations, the spleens were aseptically dissected from the immunized mice and the splenocytes then fused with mouse myeloma cells. Five thousand hybridoma clones from five cell fusions were screened for anti-WEEV and anti-EEEV NAbs respectively. As a result, five anti-WEEV and three anti-EEEV hybridoma clones were identified to secrete NAbs, which neutralized WEEV or EEEV infectivity in Vero cells. These NAbs were then produced and purified. Their neutralizing activities were confirmed in an in vitro neutralization assay. Two NAbs (G8-2-H9 and 12WA) with a 50% inhibitory concentration (IC50) of 30 ng/mL against WEEV strain 71V-1658 or two NAbs (G1-2-H4 and G1-4-C3) with IC50 of 1.5 and 4.5 μg/mL against EEEV strain PE6 were further tested against various strains of WEEV or EEEV. Both G8-2-H9 and 12WA neutralized 6 of 8 tested strains, while both G1-2-H4 and G1-4-C3 neutralized all 4 tested EEEV strains, indicating these NAbs are likely to have broadly neutralizing activity against WEEV or EEEV infectivity. To prove the concept, G8-2-H9 was evaluated in vivo and d

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vaccines;encephalitic alphaviruses;efficacy
Report Number
DRDC-RDDC-2017-R057 — Scientific Report
Date of publication
01 Apr 2017
Number of Pages
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