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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/1101
Title: Development of nsP2 protease based cell free high throughput screening assay for evaluation of inhibitors against emerging Chikungunya virus
Authors: Saha A.
Acharya B.N.
Priya R.
Tripathi N.K.
Shrivastava A.
Rao M.K.
Kesari P.
Narwal M.
Tomar S.
Bhagyawant S.S.
Parida M.
Dash P.K.
Published in: Scientific Reports
Abstract: Chikungunya virus has emerged as one of the most important global arboviral threats over the last decade. Inspite of large scale morbidity, with long lasting polyarthralgia, so far no licensed vaccine or antiviral is available. CHIKV nsP2 protease is crucial for processing of viral nonstructural polypeptide precursor to release enzymes required for viral replication, thus making it a promising drug target. In this study, high cell density cultivation (HCDC) of Escherichia coli in batch process was carried out to produce rCHIKV nsP2pro in a cost-effective manner. The purified nsP2pro and fluorogenic peptide substrate have been adapted for fluorescence resonance energy transfer (FRET) based high throughput screening (HTS) assay with Z' value and CV of 0.67 ± 0.054 and <10% respectively. We used this cell free HTS system to screen panel of metal ions and its conjugate which revealed zinc acetate as a potential candidate, which was further found to inhibit CHIKV in Vero cells. Scale-up process has not been previously reported for any of the arboviral nonstructural enzymes. The successful scale-up method for viral protease together with a HTS assay could lead to the development of industrial level large-scale screening platform for identification of protease inhibitors against emerging and re-emerging viruses. © 2018 The Author(s).
Citation: Scientific Reports(2018), 8(1): -
URI: https://doi.org/10.1038/s41598-018-29024-2
http://repository.iitr.ac.in/handle/123456789/1101
Issue Date: 2018
Publisher: Nature Publishing Group
ISSN: 20452322
Author Scopus IDs: 57102617500
36939736600
54884806200
14631231300
23978922200
55345221300
56604951200
49461618600
57203506001
13407122400
7003305014
7102314180
Author Affiliations: Saha, A., Virology Division, Defence Research and Development Establishment, Gwalior, 474002, India
Acharya, B.N., Synthetic Chemistry Division, Defence Research and Development Establishment, Gwalior, 474002, India
Priya, R., Virology Division, Defen
Funding Details: Amrita Saha is a recipient of DRDO Senior Research Fellowship. The authors would like to thank Director, DRDE for his support and providing infrastructure to carry out this work. Authors are also thankful to Dr. Nandita Saxena for flow cytometric analysis
Corresponding Author: Dash, P.K.; Virology Division, Defence Research and Development EstablishmentIndia; email: pabandash@drde.drdo.in
Appears in Collections:Journal Publications [BT]

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