SMALL SCALE EXPRESSION, SOLUBILIZATION AND CHARACTERIZATION OF CHIKUNGUNYA VIRUS STRUCTURAL PROTEINS
Abstract
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 Objective: The severity and spread of Chikungunya fever in absence of effective antiviral therapy presents a serious public health threat. The present investigation aims to generate soluble and purified viral structural proteins that can be utilized to facilitate generation of reagents for development of both diagnostic and therapeutic measures.
Methods: Bacterial expression system was used for optimization of expression and solubilization of structural proteins of CHIKV (Capsid, 6K and envelope proteins 1-3 [E1, E2 and E3]) as fusions with large (GST) and small (His and Strep) tags on a single platform. Affinity chromatography was used for small scale purification of viral proteins.
Result: The effect of different tags, inducer concentrations, temperatures and duration of induction on solubilization of proteins has been optimized and small scale purification of all the structural proteins has been attempted. Utility of these solubilized proteins has been shown by analyzing the interaction of E2 with all the structural proteins using pull down assay.
Conclusion: Small scale purification of all five structural proteins and ectodomains of envelope proteins E1 and E2 has been standardized. The data and reagents generated can be utilized for large scale purification and studying CHIKV biology.
Keywords: Chikungunya virus, Protein expression, Solubilization, Purification.
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