Enhanced Detection of Infectious Pancreatic Necrosis Virus via Lateral Flow Chip and Fluorometric Biosensors Based on Self-Assembled Protein Nanoprobes
- Authors
- Chayan, Sachin G.; Kim, Dasom; Hwang, Jangsun; Choi, Yonghyun; Hong, Jong Wook; Kim, Jeongho; Lee, Min-Ho; Hwang, Mintai P.; Choi, Jonghoon
- Issue Date
- Nov-2019
- Publisher
- AMER CHEMICAL SOC
- Keywords
- magnetic bead complex; infectious pancreatic necrosis virus; human apoferritin nanoprobe; paper-based immunoassay; Ni-NTA gold nanoparticle
- Citation
- ACS SENSORS, v.4, no.11, pp.2937 - 2944
- Indexed
- SCIE
SCOPUS
- Journal Title
- ACS SENSORS
- Volume
- 4
- Number
- 11
- Start Page
- 2937
- End Page
- 2944
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/2064
- DOI
- 10.1021/acssensors.9b01362
- ISSN
- 2379-3694
- Abstract
- Salmon fish farmers face remarkable problems in fish rearing and handling due to the spread of disease by infectious pancreatic necrosis virus (IPNV). Therefore, we developed a straightforward and sensitive technique to detect IPNV-based on recombinant human apoferritin heavy chain (hAFN-H) protein nanoparticles. In this study, the 24 subunits of the hAFN-H were genetically modified to express 6xHis-tag and protein-G at their C-terminal site using Escherichia coli. We thus achieved a two-step signal amplifying strategy that utilizes a recombinant hAFN-H nanoprobe having a protein-G-binding site that targets the F-c region of monoclonal antibodies and a 6xHis-tag that actively interacts with the functionalized Ni-NTA derivatives. In this study, we report a considerable advancement in magnetic bead-based detection systems that use Ni-NTA-Atto 550, reliably exhibiting detection limits of 1.02 TCID50/mL (50% tissue culture infective dose). Additionally, we propose a lateral flow chip-based detection method that uses the hAFN-H surface functionalized with 5 nm of the Ni-NTA-nanogold complex as a nanoprobe; the limit of detection towards IPNV was 0.88 TCID50/mL. The detection of IPNV by this recombinant hAFN-H nanoprobe was linear to virus titers in the range of 10(1)-10(3) TCID50/mL.
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