title: Highly sensitive photoelectrochemical immunoassay with enhanced amplification using horseradish peroxidase induced biocatalytic precipitation on a CdS quantum dots multilayer electrode.
abstract: Herein we demonstrate the protocol of a biocatalytic precipitation (BCP)-based sandwich photoelectrochemical (PEC) horseradish peroxidase (HRP)-linked immunoassay on the basis of their synergy effect for the ultrasensitive detection of mouse IgG (antigen, Ag) as a model protein. The hybrid film consisting of oppositely charged polyelectrolytes and CdS quantum dots (QDs) is developed by the classic layer by layer (LbL) method and then employed as the photoactive antibody (Ab) immobilization matrix for the subsequent sandwich-type Ab-Ag affinity interactions. Improved sensitivity is achieved through using the bioconjugates of HRP-secondary antibodies (Ab(2)). In addition to the much enhanced steric hindrance compared with the original one, the presence of HRP would further stimulate the BCP onto the electrode surface for signal amplification, concomitant to a competitive nonproductive absorption that lowers the photocurrent intensity. As a result of the multisignal amplification in this HRP catalyzed BCP-based PEC immunoassay, it possesses excellent analytical performance. The antigen could be detected from 0.5 pg/mL to 5.0 ng/mL with a detection limit of 0.5 pg/mL.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Zhao
authors.forename: Zheng-Yuan
authors.initials: ZY
authors.lastname: Ma
authors.forename: Pei-Pei
authors.initials: PP
authors.lastname: Yu
authors.forename: Xiao-Ya
authors.initials: XY
authors.lastname: Dong
authors.forename: Jing-Juan
authors.initials: JJ
authors.lastname: Xu
authors.forename: Hong-Yuan
authors.initials: HY
authors.lastname: Chen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D055162:Biocatalysis; D015374:Biosensing Techniques / Q000295:instrumentation*; D019187:Cadmium Compounds / Q000737:chemistry*; D004563:Electrochemistry; D004566:Electrodes; D006046:Gold; D006735:Horseradish Peroxidase / Q000378:metabolism*; D007118:Immunoassay / Q000295:instrumentation*; D007074:Immunoglobulin G / Q000032:analysis; D057230:Limit of Detection; D051379:Mice; D010783:Photometry / Q000295:instrumentation*; D045663:Quantum Dots; D013440:Sulfides / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D019187:Cadmium Compounds; D007074:Immunoglobulin G; D013440:Sulfides; C034939:cadmium sulfide; D006046:Gold; D006735:Horseradish Peroxidase