Generation of fluorescent minibodies for rapid FLISA-based detection of pine wilt disease marker
- Authors
- Kim, Seon-Hyung; Jeong, Hee-Jin
- Issue Date
- Feb-2024
- Publisher
- John Wiley and Sons Ltd
- Keywords
- BxPrx; FLISA; fluorescent immunoassay; minibody; pine wilt disease
- Citation
- Journal of Chemical Technology and Biotechnology, v.99, no.2, pp 381 - 384
- Pages
- 4
- Journal Title
- Journal of Chemical Technology and Biotechnology
- Volume
- 99
- Number
- 2
- Start Page
- 381
- End Page
- 384
- URI
- https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/31862
- DOI
- 10.1002/jctb.7539
- ISSN
- 0268-2575
1097-4660
- Abstract
- BACKGROUND: Development of a rapid and accurate detection method for a pine wilt disease (PWD) biomarker is crucial to prevent the spread of this disease in forests. In this study, we produced a recombinant minibody-type antibody against Bursaphelenchus xylophilus peroxiredoxin (BxPrx), which is a protein secreted by the PWD-causing B. xylophilus pathogen. Results: We labeled the anti-BxPrx minibody using two different fluorescent dyes, Alexa Fluor (AF) 488 and AF647, and established a fluorescence-linked immunosorbent assay (FLISA) system using these probes. Both fluorescent minibodies showed limit of detection values of 10.2 and 14.6 ng for the AF488- and AF647-labeled antibodies, respectively, suggesting the usefulness of these probes for FLISA-based sensitive detection of BxPrx. Conclusion: The entire duration for performing FLISA was 3 h, and enzyme-conjugated secondary antibody incubation, washing and enzymatic reaction steps were not required, which was comparable to traditional ELISA. Both AF488- and AF647-labeled minibodies showed high sensitivity, suggesting that these probes are useful for various fluorescence measurement techniques, which typically use green or red filters. © 2023 Society of Chemical Industry (SCI). © 2023 Society of Chemical Industry (SCI).
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