Fluorobodies combine GFP fluorescence with the binding characteristics of antibodies.
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.
Zeytun
Ahmet
A
Bioscience Division, HRL-1 TA-43 MS M888, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, USA.
Jeromin
Andreas
A
Scalettar
Bethe A
BA
Waldo
Geoffrey S
GS
Bradbury
Andrew R M
AR
D000936:Antigen-Antibody Complex / Q000032:analysis*; D005456:Fluorescent Dyes / Q000737:chemistry*; D015200:Fluoroimmunoassay / Q000379:methods*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*
D000936:Antigen-Antibody Complex; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011506:Proteins; D049452:Green Fluorescent Proteins