In situ enzymatic generation of gold for ultrasensitive amperometric sandwich immunoassay of procalcitonin.

Procalcitonin (PCT) is an important indicator for bacterial inflammatory diseases, and its sensitive, accurate and rapid detection has important clinical value. On the basis of sandwich immunoassay, glucose oxidase-catalyzed gold deposition and in-situ microliter-droplet anodic stripping voltammetry (ASV) of the enzyme-generated gold directly on the immunoelectrode, the ultrasensitive electrochemical detection of PCT is achieved. A new method of the chemical dissolution of gold by an appropriately diluted aqua regia and the simultaneous cathodic preconcentration of gold on the immunoelectrode is suggested, which gives the better performance for the ASV analysis of gold than the reported one. Under optimized conditions, the ASV peak current is linear with the common logarithm of PCT concentration from 0.05 fg mL-1 to 500 ng mL-1, with a limit of detection (LOD, S/N = 3) as low as 0.04 fg mL-1. Our method has also been used for detection of PCT in serum samples with satisfactory results.

Ultrasensitive electrochemical sensing of Hg2+ based on thymine-Hg2+-thymine interaction and signal amplification of alkaline phosphatase catalyzed silver deposition.

We report an ultrasensitive electrochemical sensor for Hg2+ detection, on the basis of two Hg2+-specific oligonucleotide probes (a thiolated capture probe and a biotinated signal probe), "terminal" signal amplification of alkaline phosphatase catalyzed deposition of silver and in situ microliter-droplet anodic stripping voltammetry (ASV). The thiolated capture probe is immobilized on an Au-plated glassy carbon electrode, and the biotinated signal probe is then attached to the electrode surface through the thymine-Hg2+-thymine interaction in the presence of Hg2+. A streptavidin-alkaline phosphatase (ALP) composite is then immobilized on the electrode surface via biotin-streptavidin interaction. The immobilized ALP can catalyze the hydrolyzation of ascorbic acid 2-phosphate trisodium salt to generate ascorbic acid, which can reduce AgNO3 to form silver deposit on the modified electrode. Quantitative analysis of Hg2+ is conducted through microliter-droplet ASV of silver after its simultaneous chemical dissolution and cathodic preconcentration on the modified electrode. The sensor can linearly respond to the common logarithm of Hg2+ concentration from 0.1nM to 250µM with a limit of detection of 0.01nM (2 ppt, S/N = 3) and a sensitivity as high as 227µAdec-1.

Selective staining of CdS on ZnO biolabel for ultrasensitive sandwich-type amperometric immunoassay of human heart-type fatty-acid-binding protein and immunoglobulin G.

We report on an ultrasensitive metal-labeled amperometric immunoassay of proteins, which is based on the selective staining of nanocrystalline cadmium sulfide (CdS) on ZnO nanocrystals and in-situ microliter-droplet anodic stripping voltammetry (ASV) detection on the immunoelectrode. Briefly, antibody 1 (Ab1), bovine serum albumin (BSA), antigen and ZnO-multiwalled carbon nanotubes (MWCNTs) labeled antibody 2 (Ab2-ZnO-MWCNTs) were successively anchored on a ß-cyclodextrin-graphene sheets (CD-GS) nanocomposite modified glassy carbon electrode (GCE), forming a sandwich-type immunoelectrode (Ab2-ZnO-MWCNTs/antigen/BSA/Ab1/CD-GS/GCE). CdS was selectively grown on the catalytic ZnO surfaces through chemical reaction of Cd(NO3)2 and thioacetamide (ZnO-label/CdS-staining), due to the presence of an activated cadmium hydroxide complex on ZnO surfaces that can decompose thioacetamide. A beforehand cathodic "potential control" in air and then injection of 7µL of 0.1M aqueous HNO3 on the immunoelectrode allow dissolution of the stained CdS and simultaneous cathodic preconcentration of atomic Cd onto the electrode surface, thus the following in-situ ASV detection can be used for immunoassay with enhanced sensitivity. Under optimized conditions, human immunoglobulin G (IgG) and human heart-type fatty-acid-binding protein (FABP) are analyzed by this method with ultrahigh sensitivity, excellent selectivity and small reagent-consumption, and the limits of detection (LODs, S/N=3) are 0.4fgmL-1 for IgG and 0.3fgmL-1 for FABP (equivalent to 73 FABP molecules in the 6µL sample employed).