Fluorescent cocaine probes: a tool for the selection and engineering of therapeutic antibodies.
Cocaine is a highly addictive drug, and despite intensive efforts, effective therapies for cocaine craving and addiction remain elusive. In recent years, we and others have reported advances in anti-cocaine immunopharmacotherapy based on specific antibodies capable of sequestering the drug before it reaches the brain. In an effort to obtain high affinity therapeutic anti-cocaine antibodies, either whole IgGs or other antibody constructs, fluorescence spectroscopic techniques could provide a means of assisting selection and engineering strategies. We report the synthesis of a series of cocaine-fluorophore conjugates (GNC-F1, GNC-F2, GNC-I) and the functional evaluation of these compounds against single-chain Fv antibodies obtained via crystallographic analysis/engineering and against commercially available anti-cocaine monoclonal antibodies with a wide range of cocaine-binding affinities. From these studies, we determined that the GNC-F2 fluorophore reproduced affinity constants obtained using [(3)H]-labeled cocaine. We anticipate that the readily synthesized and nonradioactive GNC-F2 will find use both as a tool for bioimaging and in the high-throughput selection and engineering of potential therapeutic antibodies against cocaine.
Meijler
Michael M
MM
Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, BCC-582, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Kaufmann
Gunnar F
GF
Qi
Longwu
L
Mee
Jenny M
JM
Coyle
Avery R
AR
Moss
Jason A
JA
Wirsching
Peter
P
Matsushita
Masayuki
M
Janda
Kim D
KD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D001483:Base Sequence; D003042:Cocaine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000737:chemistry* / Q000276:immunology; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D015202:Protein Engineering; D013050:Spectrometry, Fluorescence
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D018796:Immunoconjugates; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D019151:Peptide Library; D003042:Cocaine