Abstract
We report the development of a competitive heterogeneous immunoassay for BSA-conjugated Salmonella antigen using a fusion protein between an anti-Salmonella single-chain antibody (scFv) and the photo-protein aequorin (AEQ). It has been demonstrated that the fusion protein (scFvLAEQ) maintains both the antibody binding affinity toward the Salmonella antigen as well as the luminescence properties of the photoprotein. Using scFvLAEQ, the competitive immunoassay gave a detection limit of 10 μgml-1 of Salmonella antigen. The assay eliminates the need for a labeled secondary antibody usually required in enzyme-linked immuosorbent assays (ELISAs), and therefore, it may find potential applications in detecting Salmonella contamination.
| Original language | English |
|---|---|
| Pages (from-to) | 255-263 |
| Number of pages | 9 |
| Journal | Analytica Chimica Acta |
| Volume | 435 |
| Issue number | 2 |
| DOIs | |
| State | Published - May 24 2001 |
Funding
This work was supported by grants from the US National Institute of Health (Grant GM 47915) to Sylvia Daunert and Department of Energy (DE FG0595ER62010) to S. Daunert. S. Daunert is a Cottrell Scholar and a Lilly Faculty Awardee. The authors would like to thank Dr. Leonidas G. Bachas for helpful discussions. We are also thankful to Dr. Roger MacKenzie and Tomoko Hirama for the BIAcore analysis.
| Funders | Funder number |
|---|---|
| National Institutes of Health (NIH) | DE FG0595ER62010, GM 47915 |
Keywords
- Aequorin
- Bioluminescence
- Immunoassay
- Salmonella detection
- Single-chain antibody
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
- Environmental Chemistry
- Spectroscopy