Transition metal-carbonyl labeling of biotin and avidin for use in solid-phase carbonyl metallo immunoassay (CMIA).
The preparation of several transition metal-carbonyl tracers of biotin and avidin is described. Multiple labeling of avidin was achieved by acylation of some of its amine-bearing residues with N-succinimidyl 4-pentynoate (dicobalt hexacarbonyl). By varying the initial amount of this complex, protein conjugates with the extent of derivatization of up to 13 were obtained. Biotin was labeled with one (eta5-cyclopentadienyl)manganese tricarbonyl moiety by reaction of biotin hydrazide, whereas multiple labeling was reached by successive conjugation of biotin and N-succinimidyl 4-pentynoate (dicobalt hexacarbonyl) or (eta5-cyclopentadienyl)iron dicarbonyl (eta1-N-maleimidato) to poly-L-lysine or fourth generation Starburst dendrimer. All the conjugates displayed a good to excellent bioaffinity toward their respective counterparts, as measured by competitive enzymatic assays.
Salmain
Michèle
M
Ecole Nationale Supérieure de Chimie de Paris, Laboratoire de Chimie et Biochimie des Complexes Moléculaires (UMR CNRS 7576), 11 rue Pierre et Marie Curie, 75231 Paris Cedex 05, France.
Fischer-Durand
Nathalie
N
Cavalier
Laure
L
Rudolf
Bogna
B
Zakrzewski
Janusz
J
Jaouen
Gérard
G
D001360:Avidin / Q000737:chemistry*; D001667:Binding, Competitive; D001710:Biotin / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008670:Metals / Q000737:chemistry*; D011863:Radioimmunoassay; D017550:Spectroscopy, Fourier Transform Infrared; D028561:Transition Elements / Q000737:chemistry*
D008670:Metals; D028561:Transition Elements; D001360:Avidin; D001710:Biotin