- Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
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Woo-Chang Chung , Kwang Yeon Hwang , Suk-Jo Kang , Jae-Ouk Kim , Moon Jung Song
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J. Microbiol. 2020;58(1):46-53. Published online November 25, 2019
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DOI: https://doi.org/10.1007/s12275-020-9384-0
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The Chikungunya virus (CHIKV) belongs to the Alphavirus
genus of Togaviridae family and contains a positive-sense
single stranded RNA genome. Infection by this virus mainly
causes sudden high fever, rashes, headache, and severe joint
pain that can last for several months or years. CHIKV, a mosquito-
borne arbovirus, is considered a re-emerging pathogen
that has become one of the most pressing global health
concerns due to a rapid increase in epidemics. Because handling
of CHIKV is restricted to Biosafety Level 3 (BSL-3) facilities,
the evaluation of prophylactic vaccines or antivirals
has been substantially hampered. In this study, we first identified
the whole structural polyprotein sequence of a CHIKV
strain isolated in South Korea (KNIH/2009/77). Phylogenetic
analysis showed that this sequence clustered within the East/
Central/South African CHIKV genotype. Using this sequence
information, we constructed a CHIKV-pseudotyped lentivirus
expressing the structural polyprotein of the Korean
CHIKV isolate (CHIKVpseudo) and dual reporter genes of
green fluorescence protein and luciferase. We then developed
a pseudovirus-based neutralization assay (PBNA) using
CHIKVpseudo. Results from this assay compared to those
from the conventional plaque reduction neutralization test
showed that our PBNA was a reliable and rapid method to
evaluate the efficacy of neutralizing antibodies. More importantly,
the neutralizing activities of human sera from CHIKVinfected
individuals were quantitated by PBNA using CHIKVpseudo.
Taken together, these results suggest that our PBNA
for CHIKV may serve as a useful and safe method for testing
the neutralizing activity of antibodies against CHIKV
in BSL-2 facilities.
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Citations
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Yi-Ju Hsu, Tram-Anh Thi Phan, Hui-Chung Lin, Der-Jiang Chiao, Szu-Cheng Kuo, Syh-Jae Lin, Rei-Lin Kuo
Biomedical Journal.2026; 49(5): 100949. CrossRef - Luciferase unleashed: Lighting the path to vaccine approval using a novel high-throughput chikungunya virus neutralization assay
Jason Mendy, Lisa Bedell, Lauren C. Tindale, Jason S. Richardson, Na Li, Yari Fontebasso, Theodore C. Pierson, Keith Gottlieb, Lo Vang, Jonas Klingström
PLOS Neglected Tropical Diseases.2026; 20(8): e0014610. CrossRef - Pseudotyped Viruses: A Useful Platform for Pre-Clinical Studies Conducted in a BSL-2 Laboratory Setting
Sofiia N. Rizatdinova, Alina E. Ershova, Irina V. Astrakhantseva
Biomolecules.2025; 15(1): 135. CrossRef - Pseudovirus-Based Neutralization Assays as Customizable and Scalable Tools for Serological Surveillance and Immune Profiling
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Hui-Chung Lin, Shu-Fen Chang, Chien-Ling Su, Huai-Chin Hu, Der-Jiang Chiao, Yu-Lin Hsu, Hsuan-ying Lu, Chang-Chi Lin, Pei-Yun Shu, Szu-Cheng Kuo
BMC Infectious Diseases.2024;[Epub] CrossRef - Development of a Novel Chikungunya Virus-Like Replicon Particle for Rapid Quantification and Screening of Neutralizing Antibodies and Antivirals
Hui-Chung Lin, Der-Jiang Chiao, Pei-Yun Shu, Hui-Tsu Lin, Chia-Chu Hsiung, Chang-Chi Lin, Szu-Cheng Kuo, Juan E. Ludert
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Scientific Reports.2022;[Epub] CrossRef - Prevalence of Malaria and Chikungunya Co-Infection in Febrile Patients: A Systematic Review and Meta-Analysis
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Tropical Medicine and Infectious Disease.2021; 6(3): 119. CrossRef
- Spectral and structural analysis of large Stokes shift fluorescent protein dKeima570
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Yongbin Xu , Kwang Yeon Hwang , Ki Hyun Nam
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J. Microbiol. 2018;56(11):822-827. Published online October 24, 2018
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DOI: https://doi.org/10.1007/s12275-018-8319-5
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The Keima family comprises large Stokes shifts fluorescent
proteins, which are useful for dual-color fluorescence crosscorrelation
spectroscopy and multicolor imaging. dKeima570
belongs to the Keima family. It has a unique chromophore
sequence composed of CYG with an emission peak at 570
nm, but its molecular properties are unclear. We report the
spectral analysis of dKeima570 and its crystal structure at
2.0 Å resolution. The dKeima570 chromophore is mainly in
the protonation state in the entire pH range. The pH-induced
non-fluorescence state was observed below pH 4.0. The crystal
structure of the dKeima570 chromophore has a cis conformation
at pH 6.5. The chromophore is surrounded by a
unique hydrogen bonding network containing a water bridge
between Glu212 and Arg194. The analysis of the dimeric
interface of dKeima570 revealed the key residues that maintain
the oligomerization of Keima family. Structural comparisons
of dKeima570 and mKeima provided insights into
the unique large Stokes shifts characteristics of the Keima
family.
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Citations
Citations to this article as recorded by - Evaluation of AlphaFold3 prediction for post-translational modification, oligomeric assembly, and quenchable metal binding of fluorescent proteins
Ki Hyun Nam
Journal of Molecular Graphics and Modelling.2026; 142: 109169. CrossRef - Fluorescent hydrogel materials for heavy metal ion detection: design strategies, sensing mechanisms, and applications
Yuan-meng Zhou, Jian Zhu
TrAC Trends in Analytical Chemistry.2026; 203: 118999. CrossRef - Photoinduced Distance‐Dependent Electron Transfer of Fluorescent Proteins
Jianshu Dong, Qian Cao
ChemPhysChem.2025;[Epub] CrossRef - Data of crystal structure of the large Stokes shift fluorescent protein tKeima
Ki Hyun Nam, Yongbin Xu
Data in Brief.2024; 57: 110919. CrossRef - Structural Analysis of the Large Stokes Shift Red Fluorescent Protein tKeima
Ki Hyun Nam, Yongbin Xu
Molecules.2024; 29(11): 2579. CrossRef - Fluorescent Protein-Based Metal Biosensors
Ki Hyun Nam
Chemosensors.2023; 11(4): 216. CrossRef - Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP
In Jung Kim, Yongbin Xu, Ki Hyun Nam
Molecules.2022; 27(9): 2922. CrossRef - Spectroscopic Analysis of Fe Ion-Induced Fluorescence Quenching of the Green Fluorescent Protein ZsGreen
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Journal of Fluorescence.2021; 31(2): 307. CrossRef - Spectroscopic and Structural Analysis of Cu2+-Induced Fluorescence Quenching of ZsYellow
In Jung Kim, Yongbin Xu, Ki Hyun Nam
Biosensors.2020; 10(3): 29. CrossRef
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