Nature Biotechnology
21, 1473 - 1479 (2003)
Published online: 9 November 2003; | doi:10.1038/nbt911
There is a Retraction (May 2004) associated with this Article.
Retraction: Fluorobodies combine GFP fluorescence with the binding
characteristics of antibodiesAhmet Zeytun1, Andreas Jeromin2, Bethe A Scalettar3, Geoffrey S Waldo1
& Andrew RM Bradbury11
Bioscience Division, HRL-1 TA-43 MS M888, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, USA. 2
Division of Neuroscience, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77030, USA. 3
Department of Physics, Lewis and Clark College, 231 Olin Center, Portland, Oregon 97219, USA.
Correspondence should be addressed to Geoffrey S Waldo waldo@telomere.lanl.gov or Andrew RM Bradbury amb@lanl.govThe 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.
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