Development and characterization of monoclonal antibody against non-structural protein-2 of Chikungunya virus and its application
Autor(es): Chattopadhyay Soma, Kumar Abhishek, Mamidi Prabhudutta, Nayak Tapas Kumar, Das Indrani, Chhatai Jagamohan, Basantray Itishree, Bramha Umarani, Maiti Prasanta Kumar, Singh Sujay, Suryawanshi Amol Ratnakar, Chattopadhyay Subhasis
Resumo: The recent epidemics of Chikungunya viruses (CHIKV) with unprecedented magnitude - unusual clinical severity have raised a great public health concern worldwide, especially due to unavailability of vaccine or specific therapy. This emphasizes the need to underst- the biological processes of this virus in details. Although CHIKV associated research has been initiated, the availability of CHIKV specific reagents for in-depth investigation of viral infection - replication are scanty. For Alphavirus replication, non-structural protein 2 (nsP2) is known to play a key regulatory role among all other non-structural proteins. The current study describes the development - characterization of nsP2 specific monoclonal antibody (mAb) against a synthetic peptide of CHIKV. Reactivity - efficacy of this mAb have been demonstrated by ELISA, Western blot, Flow cytometry - Immunofluorescence assay. Time kinetic study confirms that this mAb is highly sensitive to CHIKV-nsP2 as this protein has been detected very early during viral replication in infected cells. Homology analysis of the selected epitope sequence reveals that it is conserved among all the CHIKV strains of different genotypes, while analysis with other Alphavirus sequences shows that none of them are 100% identical to the epitope sequence. Moreover, using the mAb, three isoforms of CHIKV-nsP2 have been detected in 2D blot analysis during infection in mammalian cells. Accordingly, it can be suggested that the mAb reported in this study can be a sensitive - specific tool for experimental investigations of CHIKV replication - infection.
Palavras-Chave: Chikungunya virus; Monoclonal antibody; nsP2
Imprenta: Journal of Virological Methods, v. 199, p. 86-94, 2014
Identificador do objeto digital: 10.1016/j.jviromet.2014.01.008
Descritores: Chikungunya virus - Cell ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Antibodies ; Chikungunya Virus - Virus ; Chikungunya virus - Vaccine ; Chikungunya virus - Epidemic ; Chikungunya virus - Immunology ; Chikungunya virus - Public health
Data de publicação: 2014
