A sulfhydryl-reactive ruthenium (II) complex and its conjugation to protein G as a universal reagent for fluorescent immunoassays

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题名:	A sulfhydryl-reactive ruthenium (II) complex and its conjugation to protein G as a universal reagent for fluorescent immunoassays
作者:	林子超;Lin, Jing-Tang;Chen, Po-Chung;Goud, Thirumani Venkatshwar;Huang, Bor-Rong;Lin, Tzu-Chau;Biellmann, Jean-François;Chen, Chien-Sheng
贡献者:	理學院化學學系
关键词:	Fluorescence;Histidine;Immunoglobulin G;Platinum group compounds;Protein binding;Reagents;Ruthenium;Sulfates;Thiols
日期:	2012-04-26
上传时间:	2026-04-21 13:55:25 (UTC+8)
出版者:	Public Library of Science
摘要:	摘要： 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. 出版者： Public Library of Science 出版日期： 2012-04-26 出處： PloS one, 2012-04, Vol.7 (4), p.e36086 資源來源： Agricultural & Environmental Science Collection 版權： COPYRIGHT 2012 Public Library of Science 識別號： ISSN: 1932-6203 識別號： EISSN: 1932-6203 識別號： DOI: 10.1371/journal.pone.0036086
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