Creating Highly Amplified ELISA Signals from Genetically Engineered Bacteriophage
- Authors
- Brasino, Michael D.; Lee, Ju Hun; Cha, Jennifer N.
- Issue Date
- Feb-2015
- Publisher
- Academic Press
- Keywords
- Bacteriophage; Biosensor; ELISA
- Citation
- Analytical Biochemistry, v.470, pp.7 - 13
- Indexed
- SCIE
SCOPUS
- Journal Title
- Analytical Biochemistry
- Volume
- 470
- Start Page
- 7
- End Page
- 13
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/18846
- DOI
- 10.1016/j.ab.2014.10.006
- ISSN
- 0003-2697
- Abstract
- For early detection of many diseases, it is critical to be able to diagnose small amounts of biomarkers in
blood or serum. One of the most widely used sensing assays is the enzyme-linked immunosorbent assay
(ELISA), which typically uses detection monoclonal antibodies conjugated to enzymes to produce colorimetric signals. To increase the overall sensitivities of these sensors, we demonstrate the use of a dually
modified version of filamentous bacteriophage Fd that produces significantly higher colorimetric signals
in ELISAs than what can be achieved using antibodies alone. Because only a few proteins at the tip of the
micron-long bacteriophage are involved in antigen binding, the approximately 4000 other coat proteins
can be augmented—by either chemical functionalization or genetic engineering—with hundreds to thousands of functional groups. In this article, we demonstrate the use of bacteriophage that bear a large
genomic fusion that allows them to bind specific antibodies on coat protein 3 (p3) and multiple biotin
groups on coat protein 8 (p8) to bind to avidin-conjugated enzymes. In direct ELISAs, the anti-rTNFa
(recombinant human tumor necrosis factor alpha)-conjugated bacteriophage show approximately 3- to
4-fold gains in signal over that of anti-rTNFa, demonstrating their use as a platform for highly sensitive
protein detection.
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