Synthesis of metal-carbonyl-dendrimer-antibody immunoconjugates: towards a new format for carbonyl metallo immunoassay.
We report the preparation of metal-carbonyl-dendrimer-antibody conjugates. These metal-carbonyl-multilabeled antibodies are designed to be used in a new solid-phase-format carbonyl metallo immunoassay (CMIA). A fourth-generation polyamidoamine dendrimer was labeled with 10-25 (eta5-cyclopentadienyl)iron dicarbonyl (eta1-N-succinimidyl) entities. An antibody was chemically modified at its carbohydrate chains by a site-directed process used to preserve the antigen-antibody binding site. The antibody was then coupled with the dendrimer labeled with 10 metal carbonyl groups. An average of 1.4 labeled dendrimers were grafted per antibody molecule. These metal-carbonyl-dendrimer-antibody conjugates were used as new universal detection reagents that recognize their specific antigens. The antigens were spotted onto nitrocellulose membranes and detected by using the conjugates in combination with Fourier transform infrared spectroscopy. A detection level in the range 5-200 pmol per membrane was achieved. This approach opens the way to a new CMIA format.
Fischer-Durand
Nathalie
N
Laboratoire de Chimie Organométallique, UMR CNRS 7576, Ecole Nationale Supérieure de Chimie de Paris, 11 rue Pierre et Marie Curie, 75231 Paris 05, France.
Salmain
Michèle
M
Rudolf
Bogna
B
Vessières
Anne
A
Zakrzewski
Janusz
J
Jaouen
Gérard
G
D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology*; D001666:Binding Sites, Antibody; D002853:Chromatography, Liquid; D003101:Collodion; D006041:Goats; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods*; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology*; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D008670:Metals / Q000737:chemistry*; D009942:Organometallic Compounds / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared
D000906:Antibodies; D018796:Immunoconjugates; D007074:Immunoglobulin G; D008670:Metals; D009942:Organometallic Compounds; D011108:Polymers; D003101:Collodion