a sulfhydryl-reactive ruthenium (ii) complex and its conjugation to protein g as a universal reagent for fluorescent immunoassayssulfhydryl-reactive钌(ii)复杂及其结合蛋白g作为荧光免疫测定的通用试剂.pdfVIP

A Sulfhydryl-Reactive Ruthenium (II) Complex and Its Conjugation to Protein G as a Universal Reagent for Fluorescent Immunoassays 1. 1. 2 3 3 Jing-Tang Lin , Po-Chung Chen , Thirumani Venkatshwar Goud , Bor-Rong Huang , Tzu-Chau Lin , 1 1 Jean-Franc¸ois Biellmann *, Chien-Sheng Chen * 1 Graduate Institute of Systems Biology and Bioinformatics, National Central University, Jhongli City, Taoyuan Country, Taiwan, 2 Institute of Chemistry, Academia Sinica, Nankang, Taipei, Taiwan, 3 Department of Chemistry, National Central University, Jhongli City, Taoyuan Country, Taiwan Abstract To develop a fluorescent ruthenium complex for biosensing, we synthesized a novel sulfhydryl-reactive compound, 4- bromophenanthroline bis-2,29-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). 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 co