A Sulfhydryl-Reactive Ruthenium (II) Complex and Its Conjugation to Protein G as a Universal Reagent for Fluorescent Immunoassays

To develop a fluorescent ruthenium complex for biosensing, we synthesized a novel sulfhydryl-reactive compound, 4-bromophenanthroline bis-2,2′-dipyridine Ruthenium bis (hexafluorophosphate). The synthesized Ru(II) complex was crosslinked with thiol-modified protein G to form a universal reagent for fluorescent immunoassays. The resulting Ru(II)-protein G conjugates were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The emission peak wavelength of the Ru(II)-protein G conjugate was 602 nm at the excitation of 452 nm which is similar to the spectra of the Ru(II) complex, indicating that Ru(II)-protein G conjugates still remain the same fluorescence after conjugation. To test the usefulness of the conjugate for biosensing, immunoglobulin G (IgG) binding assay was conducted. The result showed that Ru(II)-protein G conjugates were capable of binding IgG and the more cross-linkers to modify protein G, the higher conjugation efficiency. To demonstrate the feasibility of Ru(II)-protein G conjugates for fluorescent immunoassays, the detection of recombinant histidine-tagged protein using the conjugates and anti-histidine antibody was developed. The results showed that the histidine-tagged protein was successfully detected with dose-response, indicating that Ru(II)-protein G conjugate is a useful universal fluorescent reagent for quantitative immunoassays

SYNTHESE D'ANALOGUES DE NUCLEOSIDES ET DE NUCLEOTIDES EN VUE D'INHIBER LES POLYMERASES ET D'ETUDIER LE MECANISME DE LA RIBONUCLEOTIDE REDUCTASE DE CLASSE I

STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

SYNTHESE DE NUCLEOSIDES MODIFIES EN POSITION C-5 ET C-3 DU RIBOSE

STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

Nouveau test d'activité des NAD+glycohydrolases (activité catalytique des enzymes cristallisées)

STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF