Finely-Tuning Chemiluminescent Color of CdTe Nanocrystals and Its Application for Near-Infrared Semi-Automatic Immunoassay.

Chemiluminescence (CL), especially commercialized CL immunoassay (CLIA), is normally performed within the eye-visible region of the spectrum by exploiting the electronic-transition-related emission of the molecule luminophore. Herein, dual-stabilizers-capped CdTe nanocrystals (NCs) is employed as a model of nanoparticulated luminophore to finely tune the CL color with superior color purity. Initialized by oxidizing the CdTe NCs with potassium periodate (KIO4), intermediates of the reactive oxygen species (ROS) tend to charge CdTe NCs in both series-connection and parallel-connection routes and dominate the charge-transfer CL of CdTe NCs. The CdTe NCs/KIO4 system can exhibit color-tunable CL with the maximum emission wavelength shifted from 694 nm to 801 nm, and the red-shift span is over 100 nm. Both PL and CL of each of the CdTe NCs are bandgap-engineered; the change in the NCs surface state via CL reaction enables CL of each of the CdTe NCs to be red-shifted for ∼20 nm to PL, while the change in the NCs surface state via labeling CdTe NCs to secondary-antibody (Ab2) enables CL of the CdTe NCs-Ab2 conjugates to be red-shifted for another ∼20 nm to bare CdTe NCs. The CL of CdTe753-Ab2/KIO4 is ∼791 nm, which can perform near-infrared CL immunoassay and semi-automatically determined procalcitonin (PCT) on commercialized in vitro diagnosis (IVD) instruments.

Low-Triggering-Potential and Narrow-Potential-Window Electrochemiluminescence of Silver Nanoclusters for Gene Assay.

A low-triggering potential and a narrow-potential window are anticipated to decrease the electrochemical interference and cross talk of electrochemiluminescence (ECL). Herein, by exploiting the low oxidative potential (0.82 V vs Ag/AgCl) of dihydrolipoic acid-capped sliver nanoclusters (DHLA-AgNCs), a coreactant ECL system of DHLA-AgNCs/hydrazine (N2H4) is proposed to achieve efficient and oxidative-reduction ECL with a low-triggering potential of 0.82 V (vs Ag/AgCl) and a narrow-potential window of 0.22 V. The low-triggering-potential and narrow-potential-window nature of ECL can be primarily preserved upon labeling DHLA-AgNCs to probe DNA and immobilizing DHLA-AgNCs onto the Au surface via sandwiched hybridization, which eventually enables a selective ECL strategy for the gene assay at +0.82 V. This gene assay strategy can sensitively determine the gene of human papillomavirus from 10 to 1000 pM with a low limit of detection of 5 pM (S/N = 3) and would open a way to improve the applied ECL bioassay.

Potential-selective electrochemiluminescence of AgInS2/ZnS nanocrystals and its immunoassay application.

Potential-selective electrochemiluminescence (ECL) with tunable maximum-emission-potential ranging from 0.95 to 0.30 V is achieved using AgInS2/ZnS nanocrystals, which is promising in the design of multiplexed bioassay on commercialized ECL setups. The model system AgInS2/ZnS/N2H4 exhibits efficient ECL around 0.30 V and can be exploited for sensitive immunoassays with less electrochemical interference and crosstalk.

Silver Nanocluster-Tagged Electrochemiluminescence Immunoassay with a Sole and Narrow Triggering Potential Window.

The commercialized electrochemiluminescence (ECL) immunoassay is carried out by holding luminophore Ru(bpy)32+ at a given potential. Designing an electrochemiluminophore with a narrow triggering potential window is strongly anticipated to decrease the electrochemical cross-talk and improve the flux of the commercialized ECL immunoassay in a potential-resolved way. Herein, L-penicillamine-capped silver nanoclusters (LPA-AgNCs) are facilely synthesized and utilized as tags to perform the ECL immunoassay with a sole and narrow triggering potential window of 0.24 V by employing hydrazine (N2H4) as a coreactant. The maximum ECL emission of the LPA-AgNCs/N2H4 system is located ca. +1.27 V. Upon immobilizing LPA-AgNCs onto the electrode surface via forming a sandwich immunocomplex, the ECL of LPA-AgNCs/N2H4 can be utilized to sensitively and selectively determine human carcinoembryonic antigen from 0.5 to 1000 pg/mL with a low limit of detection of 0.1 pg/mL (S/N = 3). This work might open a way to screen electrochemiluminophores for the multiple ECL immunoassay in a potential-resolved way.

Dual-potential encoded electrochemiluminescence for multiplexed gene assay with one luminophore as tag.

Multiplexed gene assay for simultaneously detecting the multi-targets of nucleic acids is strongly anticipated for the accurate diseases diagnosis and prediction, and all commercial available gene assays for IVD are a kind of single-target assay. Herein, a dual-potential encoded and coreactant-free electrochemiluminescence (ECL) strategy is proposed for the multiplexed gene assay, which can be conveniently carried out by directly oxidizing the same luminescent tag of dual-stabilizers-capped CdTe nanocrystals (NCs). The CdTe NCs linked with sulfhydryl-RNA via Cd-S bond merely exhibits one ECL process around 0.32 V with a narrow triggering-potential-window of 0.35 V, while CdTe NCs linked with amino-RNA via amide linkage solely gives off one ECL process around 0.82 V with a narrow triggering-potential-window of 0.30 V. Multiplexing ECL of both sulfhydryl-RNA-functionalized CdTe NCs and amino-RNA-functionalized CdTe NCs can be utilized to simultaneously detect the open reading frame 1ab (ORF1ab) and the nucleoprotein (N) genes without crosstalk, in which ECL of sulfhydryl-RNA-functionalized CdTe NCs can dynamically determine ORF1ab from 200 aM to 10 fM with a limit of detection (LOD) of 100 aM, while ECL of amino-RNA-functionalized CdTe NCs can linearly detect N gene from 5 fM to 1 pM with a LOD of 2 fM. Post-engineering CdTe NCs with RNA in a labeling-bond engineering way would provide a potential-selective and encoded ECL strategy for multiplexed gene assay with one luminophore.

Luminophore-Surface-Engineering-Enabled Low-Triggering-Potential and Coreactant-Free Electrochemiluminescence for Protein Determination.

Coreactant-free electrochemiluminescence (ECL) is promising for removing the exogenous effects of coreactant and simplify the operation procedures and setups of commercialized ECL bioassays. Herein, an electrosterically involved strategy for achieving a low-triggering-potential (+0.21 V vs Ag/AgCl) and coreactant-free ECL from dual-stabilizer-capped CdTe nanocrystals (NCs) is proposed with mercaptopropionic acid (MPA) and hexametaphosphate (HMP) as the capping agents of luminophores. Upon employing the CdTe NCs as the ECL tag for the immunoassay, all the tags in the bioconjugates of the CdTe NCs and the secondary antibody (Ab2|CdTe) as well as in the final achieved sandwich-type immunocomplexes can exhibit efficient coreactant-free ECL with an electrosterically involved triggering potential nature. The bioconjugates of Ab2|CdTe with Ab2 no more than 30 kDa, such as the thyroid stimulating hormone (30 kDa) and the recombinant pro-gastrin releasing peptide (ProGRP, 14 kDa), merely exhibit coreactant-free ECL around +0.24 V, while bioconjugates of Ab2|CdTe with an Ab2 beyond 30 kDa only give off coreactant-free ECL around +0.82 V. Due to the further enhanced electrosteric effect in sandwich-type immunocomplexes, only the ECL immunosensor with ProGRP as the target can give off coreactant-free ECL around +0.24 V. The electrosterically involved and coreactant-free ECL of CdTe NCs is consequently utilized to sensitively and selectively determine the molecular protein ProGRP, which demonstrates a wide linearity range from 0.1 to 2000 pg/mL and a low limit of detection at 0.05 pg/mL (S/N = 3). This low-triggering-potential and coreactant-free combined ECL platform indicates that engineering the surface of CdTe NCs with a protein can improve the performance of ECL tags in a protein-weight-involved electrosterical way.