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High yield expression and purification of Chikungunya virus E2 recombinant protein and its evaluation for serodiagnosis
- Source :
- Journal of virological methods. 235
- Publication Year :
- 2016
-
Abstract
- Disease caused by Chikungunya virus (CHIKV) is clinically characterized by sudden-onset of fever and severe arthralgia, which may persist for weeks, months, or years after acute phase of the infection. CHIKV is spreading globally; in India it first appeared in the 1960s followed by a quiescent period and then a full-blown remergence in 2006 and sporadic persistence since then. Despite a large number of commercially available diagnostic kits for CHIKV, clinical preparedness and diagnostics suffer from sub-optimal assays. An international diagnostic laboratory survey suggested that there is a critical need for improved CHIKV diagnostics especially in the early acute phase of illness. With the recent studies indicating that a vast majority of human humoral response in CHIKV infection is directed against E2 protein, this supports strong interest to develop CHIKV E2 based serological tests. However, methods to produce large amounts of CHIKV protein are limited. Here we report cloning, expression and purification methods for obtaining a truncated 37kDa Chikungunya E2 protein at a high yield of 65-70mg/l. We found that this purified protein can be reliably used in ELISA and western blot to detect CHIKV specific antibodies in sera from patients who were PCR or IgM positive. Thus, using this protocol, laboratories can make large quantities of purified protein that can be potentially used in CHIKV serological analysis.
- Subjects :
- 0301 basic medicine
Protein Folding
Enzyme-Linked Immunosorbent Assay
Disease
medicine.disease_cause
Antibodies, Viral
Sensitivity and Specificity
Virus
Immunoglobulin G
Chromatography, Affinity
Serology
law.invention
03 medical and health sciences
Western blot
Viral Envelope Proteins
law
Virology
medicine
Humans
Chikungunya
Antigens, Viral
medicine.diagnostic_test
biology
virus diseases
Recombinant Proteins
030104 developmental biology
Immunoglobulin M
biology.protein
Recombinant DNA
Chikungunya Fever
Chikungunya virus
Subjects
Details
- ISSN :
- 18790984
- Volume :
- 235
- Database :
- OpenAIRE
- Journal :
- Journal of virological methods
- Accession number :
- edsair.doi.dedup.....8ab8c1668c6229dcb063b1d0f35f66e4