Development and characterization of monoclonal antibody against non-structural protein-2 of Chikungunya virus and its application.
The recent epidemics of Chikungunya viruses (CHIKV) with unprecedented magnitude and 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 understand 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 and 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 and characterization of nsP2 specific monoclonal antibody (mAb) against a synthetic peptide of CHIKV. Reactivity and efficacy of this mAb have been demonstrated by ELISA, Western blot, Flow cytometry and 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 and specific tool for experimental investigations of CHIKV replication and infection.
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India. Electronic address: sochat.ils@gmail.com.
Soma
S
Chattopadhyay
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Abhishek
A
Kumar
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Prabhudutta
P
Mamidi
School of Biological Sciences, National Institute of Science Education & Research, Bhubaneswar, India.
Tapas Kumar
TK
Nayak
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Indrani
I
Das
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Jagamohan
J
Chhatai
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Itishree
I
Basantray
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Umarani
U
Bramha
Imgenex India Pvt. Ltd., Bhubaneswar, India.
Prasanta Kumar
PK
Maiti
Imgenex India Pvt. Ltd., Bhubaneswar, India.
Sujay
S
Singh
Institute of Life Sciences, Nalco Square, Bhubaneswar 751023, India.
Amol Ratnakar
AR
Suryawanshi
School of Biological Sciences, National Institute of Science Education & Research, Bhubaneswar, India.
Subhasis
S
Chattopadhyay
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000302:isolation & purification*; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western; D002646:Chikungunya virus / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000032:analysis* / Q000276:immunology; D014776:Virus Cultivation
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D017361:Viral Nonstructural Proteins
Chikungunya virus; Monoclonal antibody; nsP2