We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Fluorobodies combine GFP fluorescence with the binding characteristics of antibodies.
- Authors
Zeytun, Ahmet; Jeromin, Andreas; Scalettar, Bethe A.; Waldo, Geoffrey S.; Bradbury, Andrew R. M.
- Abstract
The difficulty of deriving binding ligands to targets identified by genomic sequencing has led to a bottleneck in genomic research. By inserting diverse antibody binding loops into four of the exposed loops at one end of green fluorescent protein (GFP), we have mimicked the natural antibody binding footprint to create robust binding ligands that combine the advantages of antibodies (high affinity and specificity) with those of GFP (intrinsic fluorescence, high stability, expression and solubility). These 'fluorobodies' have been used effectively in enzyme-linked immunosorbent assays (ELISAs), flow cytometry, immuno-fluorescence, arrays and gel shift assays, and show affinities as high as antibodies. Furthermore, the intrinsic fluorescence of fluorobodies correlates with binding activity, allowing the rapid determination of functionality, concentration and affinity. These properties render them especially suitable for the high-throughput genomic scale selections required in proteomics, as well as in diagnostics, target validation and drug development.
- Subjects
NUCLEOTIDE sequence; GREEN fluorescent protein; GENETIC research; LIGAND binding (Biochemistry); ENZYME-linked immunosorbent assay; IMMUNOFLUORESCENCE; IMMUNOGLOBULINS; DRUG development
- Publication
Nature Biotechnology, 2003, Vol 21, Issue 12, p1473
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt911