Fluorobodies combine GFP fluorescence with the binding characteristics of antibodies
Just parking, but looks very interesting
Ahmet Zeytun1, Andreas Jeromin2, Bethe A Scalettar3, Geoffrey S Waldo1 & Andrew RM Bradbury1
1. 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 G S Waldo. e-mail: waldo@telomere.lanl.gov and A R M Bradbury. e-mail: amb@lanl.gov
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.
Published online: 9 November 2003, doi:10.1038/nbt911
December 2003 Volume 21 Number 12 pp 1473 - 1479
and the patent applicaton
United States Patent Application 20030203355
Kind Code A1
Bradbury, Andrew M. ; et al. October 30, 2003
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Fluorobodies: binding ligands with intrinsic fluorescence
Abstract
The current invention provides binding ligands with intrinsic fluorescent activity. The invention also provides libraries of such binding ligands, methods of preparing such binding ligands and libraries, and methods of identifying a binding ligand that specifically binds to a target molecule.
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Inventors: Bradbury, Andrew M.; (Santa Fe, NM) ; Zeytun, Ahmet; (Los Alamos, NM) ; Waldo, Geoffrey S.; (Santa Fe, NM)
Correspondence Name and Address:
TOWNSEND AND TOWNSEND AND CREW, LLP
TWO EMBARCADERO CENTER
EIGHTH FLOOR
SAN FRANCISCO
CA
94111-3834
US
Assignee Name and Adress: Los Alamos National Laboratory
1111 Franklin Street, 12th Floor
Los Alamos
NM
94607
The Regents of the University of California
Oakland
CA
Serial No.: 132067
Series Code: 10
Filed: April 24, 2002
U.S. Current Class: 435/5; 435/6; 435/7.1; 530/350
U.S. Class at Publication: 435/5; 435/6; 435/7.1; 530/350
Intern'l Class: C12Q 001/70; C12Q 001/68; G01N 033/53; C07K 014/435 |
