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Fig. 2 | Virology Journal

Fig. 2

From: Optimization of Zika virus envelope protein production for ELISA and correlation of antibody titers with virus neutralization in Mexican patients from an arbovirus endemic region

Fig. 2

Characterisation of ZIKV Env proteins. a. Silver stained PAGE gel showing purity of ZIKV Env proteins: commercial Env based on African Strain (left panel), Env-CD4 (middle) and Env (right) based on Asian-lineage strain. Three concentrations of Env proteins were used: 500 ng, 250 ng and 125 ng for African ZIKV Env (commercial) and Asian-lineage ZIKV Env-CD4. Env-CD4 showed three distinctive bands at ~ 65 kDa, ~ 45 kDa and ~ 25 kDa on the gel, corresponding to Env-CD4, Env and CD4 fusion tag (CD4), respectively. CD4 fragment is liberated following cleavage from Env-CD4 protein into Env and CD4. b. Western blot of the purified ZIKV Env-CD4 and Env proteins using a pan-flavivirus antibody, sera from ChAdOx1 ZIKV prME_ΔTM immunized mice and Zika Env monoclonal antibody. For Env-CD4, the top band corresponds to Env-CD4 protein and the bottom band to Env protein which may be formed following cleavage from Env-CD4. c. Western blot of the purified Env-CD4 using an anti-His antibody (1:2000). For Env-CD4, the top band is Env-CD4 protein and bottom band corresponds to CD4 (His). An unrelated malaria protein (cCSP-His) was used as positive control for the His antibody and as a negative control for the ZIKV-Env specific antibodies. Negative control is cell-free media

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