Abstract:
:Arthropod-borne flaviviruses such as dengue virus (DENV) and West Nile virus (WNV) pose significant health threats to the global community. Due to escalating numbers of DENV and WNV infections worldwide, development of an effective vaccine remains a global health priority. As flavivirus envelope Domain III (DIII) protein is highly immunogenic and capable of inducing neutralizing antibodies against wild-type virus, it is both a potential protein subunit vaccine candidate and a suitable diagnostic reagent. Here, we describe the use of metal affinity membrane chromatography as a rapid and improved alternative for the purification of recombinant DIII (rDIII) antigens from DENV serotypes 1-4 and WNV - New York, Sarafend, Wengler and Kunjin strains. Optimum conditions for the expression, solubilization, renaturation and purification of these proteins were established. The purified proteins were confirmed by MALDI-TOF mass spectrometry and ELISA using antibodies raised against the respective viruses. Biological function of the purified rDIII proteins was confirmed by their ability to generate DIII-specific antibodies in mice that could neutralize the virus.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Tan LC,Chua AJ,Goh LS,Pua SM,Cheong YK,Ng MLdoi
10.1016/j.pep.2010.06.015subject
Has Abstractpub_date
2010-11-01 00:00:00pages
129-37issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(10)00190-7journal_volume
74pub_type
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