Development of a Spacer-optimized Quenchbody against Tumor Necrosis Factor AlphaDevelopment of a Spacer-optimized Quenchbody against Tumor Necrosis Factor Alpha
- Other Titles
- Development of a Spacer-optimized Quenchbody against Tumor Necrosis Factor Alpha
- Authors
- 윤한울; Hiroshi Ueda; 정희진
- Issue Date
- 1-Oct-2022
- Publisher
- 한국생물공학회
- Keywords
- antibody fragment; Quenchbody; fluorescence immunosensor; tumor necrosis factor alpha; on-site immunoassay
- Citation
- Biotechnology and Bioprocess Engineering, v.27, no.5, pp.820 - 830
- Journal Title
- Biotechnology and Bioprocess Engineering
- Volume
- 27
- Number
- 5
- Start Page
- 820
- End Page
- 830
- URI
- https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/30565
- DOI
- 10.1007/s12257-022-0088-7
- ISSN
- 1226-8372
- Abstract
- Tumor necrosis factor alpha (TNFα) is a proinflammatory cytokine, and its overproduction causes infectious diseases and autoimmune disorders. Therefore, rapid and accurate detection of TNFα is important, and the need for a convenient detection system has increased. In this study, single-chain variable fragment (scFv)-type Quenchbody (Q-body) was developed that could detect TNFα by mixing the Q-body with TNFα, followed by measuring its fluorescence intensity. Recombinant scFvs were constructed with several different lengths of flexible GGGS repeat spacers between dye and scFv. The fluorescent responses of Q-bodies increased in an antigen-dosedependent manner. The fluorescence intensity in the presence of a maximum concentration of antigen was compared to that of the denatured Q-body, resulting in the Q-body with the (GGGS)5 spacer showing a fully de-quenched signal, whereas other Q-bodies with shorter spacer lengths showed insufficient de-quenching efficiency. These results indicated that adjusting the spacer length is important for improving the response of the Q-body. The Q-body with the optimized spacer length showed a maximum 4.5-fold increase in fluorescence intensity with a broad antigen concentration range, and nanomolar order of detection limit, indicating the usefulness of this Q-body for one-step detection of TNFα.
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Collections - College of Science and Technology > Department of Biological and Chemical Engineering > 1. Journal Articles
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