Liu, Jinny L.Webb, Emily M.Zabetakis, DanBurke, Crystal W.Gardner, Christina L.Glass, Pamela J.Legler, Patricia M.Weger-Lucarelli, JamesAnderson, George P.Goldman, Ellen R.2021-05-282021-05-282021-01-28626028http://hdl.handle.net/10919/103547A single domain antibody (clone CC3) previously found to neutralize a vaccine strain of the chikungunya virus (PRNT50 = 2. 5 ng/mL) was found to be broadly neutralizing. Clone CC3 is not only able to neutralize a wild-type (WT) strain of chikungunya virus (CHIKV), but also neutralizes WT strains of Mayaro virus (MAYV) and Ross River virus (RRV); both arthralgic, Old World alphaviruses. Interestingly, CC3 also demonstrated a degree of neutralizing activity against the New World alphavirus, Venezuelan equine encephalitis virus (VEEV); albeit both the vaccine strain, TC-83, and the parental, WT Trinidad donkey strain had PRNT50 values similar to 1,000-fold higher than that of CHIKV. However, no neutralization activity was observed with Western equine encephalitis virus (WEEV). Ten CC3 variants designed to possess a range of isoelectric points, both higher and lower, were constructed. This approach successfully identified several lower pI mutants which possessed improved thermal stabilities by as much as 10 degrees C over the original CC3 (T-m = 62 degrees C), and excellent refolding abilities while maintaining their capacity to bind and neutralize CHIKV.application/pdfenCreative Commons Attribution 4.0 Internationalchikungunya virusold worldnew worldalphavirusneutralizationmelting temperaturesingle domain antibodyArticle - Refereedhttps://doi.org/10.3389/fmed.2021.6260288335855272296-858X