Yingshuai Liu

Aptamer induced assembly of fluorescent nitrogen-doped carbon dots on gold nanoparticles for sensitive detection of AFB1.

Novel fluorescent nitrogen-doped carbon dots (N,C-dots) were synthesized and assembled on aptamer modified gold nanoparticles (Aptamer/AuNPs) for the super sensitive detection of aflatoxin B1 (AFB1). Positively charged N,C-dots were synthesized by the hydrothermal treatment of pancreatin. The prepared N,C-dots were assembled on aptamer/AuNPs by electrostatic interactions. The fluorescence of the N,C-dots was efficiently quenched. When AFB1 was added to the assay solution, specific interactions between AFB1 and the aptamer caused release of the N,C-dots. The fluorescence of the N,C-dots recovered and the intensity increase could be used to calculate the amount of AFB1 added. The assay exhibits super-high sensitivity with a detection limit of 5 pg/mL (16 pM) and a wide range of linear response of 5 pg/mL to 2.00 ng/mL. A novel aptasensor is thus successfully constructed, it provides an efficient way for sensitive AFB1 sensing as well as a new technique for aptamer based novel sensor construction.

ZnO nanorods-enhanced fluorescence for sensitive microarray detection of cancers in serum without additional reporter-amplification

Current high-throughput antibody microarrays greatly rely on enzymatic reactions- or nanostructured tags-based reporter-amplifications for required sensitivity, but they need tedious operation, additional expensive reagents and microwells or microchannels to eliminate crossover interferences, thus resulting in low array density and high expense. A unique ZnO nanorods-grown substrate is developed here to not only immobilize a large amount of probe molecules, but also directly amplify the microarray fluorescent signals in detection of two important cancer biomarkers, carcinoembryonic antigen (CEA) and α-fetoprotein (AFP), achieving a detection limit of 1 pg mL(-1) in human serum, which is comparative to or lower than that of ELISA. This advanced ZnO nanorods-based substrate can be mass-manufactured, which offers great potential to fabricate economical and sensitive protein arrays for broad applications in clinic diagnosis, therapeutic monitoring and drug discovery.

On-demand droplet release for droplet-based microfluidic system

On-demand droplet release from microwell was successfully implemented and well combined with droplet trapping/fusion functions to make an ideal and integrated droplet based microfluidic system.

In Situ Surface Plasmon Resonance Investigation of the Assembly Process of Multiwalled Carbon Nanotubes on an Alkanethiol Self-Assembled Monolayer for Efficient Protein Immobilization and Detection

In situ surface plasmon resonance (SPR) was used to study the assembly process of multiwalled carbon nanotubes (MWCNTs) quantitatively on an alkanethiol self-assembled monolayer (SAM) surface, showing that MWCNTs can follow the Langmuir adsorption kinetics to assemble spontaneously whereas the assembly temperature has an essential influence on the assembly kinetics and the surface distribution of MWCNTs. To further in situ investigate protein attachment on the MWCNT surface and its sensing application quantitatively, goat IgG was immobilized by three strategies: direct adsorption, covalent binding, and 1-pyrenebutanoic acid, succinimidyl ester (PBSE)-assisted attachment, of which the covalent binding approach provides the best protein loading capacity. The SPR label-free detection of anti-goat IgG demonstrates excellent performance with high sensitivity, good specificity, and rapid response in comparison to that with a plain substrate without MWCNT assembly reported in our previous work. This is contributed by the 3D MWCNT assembly matrix providing a high probe immobilization capability and superb accessibility for the target to enhance its sensing performance significantly.

Randomly oriented ZnO nanorods as advanced substrate for high-performance protein microarrays.

When aligned or patterned nanostructures attract great interest, the randomly arranged nanostructures are often ignored. A nanostructure with randomly oriented ZnO nanorods was prepared on the glass slides using scalable method as a protein microarray substrate. It demonstrates significant fluorescence enhancement and superior performance over the aligned ZnO nanorods for high performance protein microarray applications.

Disposable lateral flow-through strip for smartphone-camera to quantitatively detect alkaline phosphatase activity in milk

A disposable lateral flow-through strip was developed for smartphone to fast one-step quantitatively detect alkaline phosphatase (ALP) activity in raw milk. The strip comprises two functional components, a conjugation pad loaded with phosphotyrosine-coated gold nanoparticles (AuNPs@Cys-Try-p) and a testing line coated with anti-phosphotryosine antibody (anti-Tyr-p mAb). The dephosphorylation activity of ALP at the testing zone can be quantitatively assayed by monitoring the accumulated AuNPs-induced color changes by smartphone camera, thus providing a highly convenient portable detection method. A trace amount of ALP as low as 0.1UL(-1) with a linear dynamic range of 0.1-150UL(-1) (R(2)=0.999) in pasteurized milk and raw milk can be one-step detected by the developed flow-through strip within 10min, demonstrating the potential of smartphone-based portable sensing device for pathogen detection. This bio-hazards free lateral flow-through testing strip can be also used to fabricate rapid, sensitive and inexpensive enzyme or immunosensors for broad portable clinic diagnosis and food contamination analysis, particularly in point-of-care and daily food quality inspection.

Hybrid ZnO Nanorod-Polymer Brush Hierarchically Nanostructured Substrate for Sensitive Antibody Microarrays

A hierarchically nanostructured organic-inorganic hybrid substrate comprising randomly oriented ZnO nanorods on glass slide with coaxially tethered dense polymer brush, POEGMA-co-GMA is reported for highly sensitive antibody microassay, achieving excellent detection specificity, and superior detection limit of as low as 100 fg mL(-1) for biomarkers in human serum within a 1 h assay time.

High performance protein microarrays based on glycidyl methacrylate-modified polyethylene terephthalate plastic substrate

There is a great challenge to immobilize high density of probe molecules for high performance protein microarrays, and this is achieved in this work by using polyethylene terephthalate (PET) plastic substrate onto which glycidyl methacrylate (GMA) photopolymer is grafted under mild conditions to introduce high density of epoxy groups for covalent immobilization of proteins. The poly(GMA)-grafted PET (PGMA-PET) surface was characterized with atomic force microscope (AFM) and attenuated total reflectance Fourier transform infra-red (ATR-FTIR) spectroscopy. For high density of protein immobilization and good quality of microspots, experiments were conducted to optimize the printing buffer, and an optimal buffer was found out to be PBS with 10% glycerol+0.003% triton X-100. According to the studies of loading capacity and immobilization kinetics, the optimal protein probe concentration and incubation time for the efficient immobilization are 200 microg mL(-1) and 8h, respectively. The performance of the PGMA-PET-based protein microarrays is evaluated with sandwich immunoassay using rat IgG and anti-rat IgG as model proteins, demonstrating a limit of detection (LOD) of 10 pg mL(-1) and a dynamic range of five orders of magnitude which are better than or very comparable with the reported or commercially available immunoassays, while providing a high-throughput approach. The work renders a simple and economic method to manufacture high performance protein microarrays and is expected to have great potentials in broad applications related to clinic diagnosis, drug discovery and proteomic research.

High-performance UV-curable epoxy resin-based microarray and microfluidic immunoassay devices.

Immunoassay devices including microarray and microfluidic systems were fabricated with an UV-curable resin by a new economic approach, which can not only simply produce a 3-dimensional (3D) patterned structure, but also simultaneously introduce functional epoxide groups for efficient protein immobilization. The performance of the epoxy resin-based microarray was improved by optimization of printing buffer, probe concentration, and immobilization time, showing a detection dynamic range of 5 orders of magnitude and a limit of detection (LOD) of 10 pg mL(-1) for immunoglobulin G (IgG). The developed microfluidic immunoassay device demonstrates a LOD of 100 pg mL(-1) for IL-5 detection. The device can also be used to colorimetrically detect proteins via naked human eyes for immunoassays. This work provides a simple and inexpensive method to fabricate a sensitive immunoassay device, especially a 3D microfluidic system, which has great potential to develop a portable immunoassay device via human eye detection for point-of-care service and/or high throughput screening of infectious diseases.

Photografted poly(methyl methacrylate)-based high performance protein microarray for hepatitis B virus biomarker detection in human serum

A robust and efficient strategy is investigated to functionalize poly(methyl methacrylate) as a superior substrate for high performance protein microarrays and utilized for sensitive detection of hepatitis B virus biomarker in human serum, demonstrating its great potential applications in proteomics and clinic diagnosis.