title: Ultrasensitive and fast fluorescent bioassay based on fluorescence enhancement of silver nanoparticles.
abstract: An ultrasensitive, fast and specific fluorescent platform for protein detection is developed. In this protocol, silver nanoparticles were conjugated with paramagnetic particles (MPs-Ag) for target capture, concentration and separation; fluorescent dyes functionalized silver nanoparticles (Tag) for generating signals. The presented method is highly sensitive and specific with a detection limit of 2.2 pM for thrombin, and no significant interference was observed for other proteins such as human serum albumin (HSA), lysozyme and IgG. This novel approach combining the magnetic separation and concentration of MPs-Ag, aptamer recognition and fluorescence enhancement of Tag, can be successfully used to enhance the sensitivity of detecting ultra-low levels of target proteins or biomolecules.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, China. xudanke@nju.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.forename: Weibing
authors.initials: W
authors.lastname: Qiang
authors.forename: Chongzhi
authors.initials: C
authors.lastname: Wang
authors.forename: Maika
authors.initials: M
authors.lastname: Vuki
authors.forename: Danke
authors.initials: D
authors.lastname: Xu
mesh_terms: D001483:Base Sequence; D001681:Biological Assay / Q000379:methods*; D017931:DNA Primers; D005453:Fluorescence; D053768:Metal Nanoparticles; D046529:Microscopy, Electron, Transmission; D012680:Sensitivity and Specificity; D012834:Silver / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D017931:DNA Primers; D012834:Silver