Vertically Ordered Mesoporous Silica Film-Assisted Label-Free and Universal Electrochemiluminescence Aptasensor Platform.

Herein, a simple, facile, and label-free electrochemiluminescence (ECL) aptasensor platform was constructed by integration of aptamer-gated systems and vertically ordered mesoporous silica films (MSFs) grown in suit of indium-tin oxide (ITO) electrode. In this aptasensor platform, aptamer could be effectively adsorbed on the surface of aminated MSFs by noncovalent electrostatic attraction and then employed as an ideal gate material to control the blocking and releasing of luminescence reagents (Ru(bipy)32+) entrapped within the pores of MSFs. In the presence of target, the specific aptamer-target binding could trigger the uncapping the pores, releasing the Ru(bipy)32+ with detectable reduced of ECL signal. The feasibility and universality of this design was validated by employing three aptamers that bind to lysozyme, adenosine, and K+ as gate materials, and the detection limits were determined to be 0.06 nM, 0.75 nM, and 0.5 µM, respectively. This ECL aptasensor, based on the simple competitive procedure, was simple design, undemanding, and fast in operation. In addition, no other chemical modification of the aptamer was required, suggesting that this ECL aptasensor could be applied to many other target detections just by altering the aptamer sequence.

Self-calibrated atom-interferometer gyroscope by modulating atomic velocities.

Atom-interferometer gyroscopes have attracted much attention for their long-term stability and extremely low drift. For such high-precision instruments, self-calibration to achieve an absolute rotation measurement is critical. In this work, we propose and demonstrate the self-calibration of an atom-interferometer gyroscope. This calibration is realized by using the detuning of the laser frequency to control the atomic velocity, thus modulating the scale factor of the gyroscope. The modulation determines the order and the initial phase of the interference stripe, thus eliminating the ambiguity caused by the periodicity of the interferometric signal. This self-calibration method is validated through a measurement of the Earth's rotation rate, and a relative uncertainty of 162 ppm is achieved. Long-term stable and self-calibrated atom-interferometer gyroscopes have important applications in the fields of fundamental physics, geophysics, and long-time navigation.

Compact multi-channel radio frequency pulse-sequence generator with fast-switching capability for cold-atom interferometers.

Cold-atom interferometers have matured into a powerful tool for fundamental physics research, and they are currently moving from realizations in the laboratory to applications in the field. A radio frequency (RF) generator is an indispensable component of these devices for controlling lasers and manipulating atoms. In this work, we developed a compact RF generator for fast switching and sweeping the frequencies and amplitudes of atomic-interference pulse sequences. In this generator, multi-channel RF signals are generated using a field-programmable gate array (FPGA) to control eight direct digital synthesizers (DDSs). We further propose and demonstrate a method for pre-loading the parameters of all the RF pulse sequences to the DDS registers before their execution, which eliminates the need for data transfer between the FPGA and DDSs to change RF signals. This sharply decreases the frequency-switching time when the pulse sequences are running. Performance characterization showed that the generated RF signals achieve a 100 ns frequency-switching time and a 40 dB harmonic-rejection ratio. The generated RF pulse sequences were applied to a cold-atom-interferometer gyroscope, and the contrast of atomic interference fringes was found to reach 38%. This compact multi-channel generator with fast frequency/amplitude switching and/or sweeping capability will be beneficial for applications in field-portable atom interferometers.

Aptamer/target binding-induced triple helix forming for signal-on electrochemical biosensing.

Owing to its diversified structures, high affinity, and specificity for binding a wide range of non-nucleic acid targets, aptamer is a useful molecular recognition tool for the design of various biosensors. Herein, we report a new signal-on electrochemical biosensing platform which is based on an aptamer/target binding-induced strand displacement and triple-helix forming. The biosensing platform is composed of a signal transduction probe (STP) modified with a methylene blue (MB) and a sulfhydryl group, a triplex-forming oligonucleotides probe (TFO) and a target specific aptamer probe (Apt). Through hybridization with the TFO probe and the Apt probe, the self-assembled STP on Au electrode via Au-S bonding keeps its rigid structure. The MB on the STP is distal to the Au electrode surface. It is eT off state. Target binding releases the Apt probe and liberates the end of the MB tagged STP to fold back and form a triplex-helix structure with TFO (STP/TFO/STP), allowing MB to approach the Au electrode surface and generating measurable electrochemical signals (eT ON). As test for the feasibility and universality of this signal-on electrochemical biosensing platform, two aptamers which bind to adenosine triphosphate (ATP) and human α-thrombin (Tmb), respectively, are selected as models. The detection limit of ATP was 7.2 nM, whereas the detection limit of Tmb was 0.86 nM.

A highly sensitive electrochemiluminescence assay for protein kinase based on double-quenching of graphene quantum dots by G-quadruplex-hemin and gold nanoparticles.

A highly sensitive electrochemiluminescence (ECL) strategy was developed for the protein kinase A (PKA) activity and inhibition assay based on double-quenching of graphene quantum dots (GQDs) ECL by G-quadruplex-hemin DNAzyme and gold nanoparticles (AuNPs). In this strategy, the GQDs were modified onto the indium-tin oxide (ITO) electrode and further assembled with substrate peptide of target protein kinase through covalent coupling, which can exhibit high and stable ECL signal. The AuNPs, functionalized with the phosphorylated DNA and G-quadruplex-hemin DNAzyme via Au-S chemistry, were selected as quenching probes. In the presence of PKA, the peptide on the electrode was phosphorylated and the AuNPs functionalized with the phosphorylated DNA and G-quadruplex-hemin DNAzyme were subsequently integrated onto the phosphorylated peptide by Zr(4+). Owing to the reduction of coreactant H2O2 resulting from G-quadruplex-hemin DNAzyme catalytic reaction and the ECL energy transfer (ECL-RET) between AuNPs and GQDs, the ECL intensity of GQDs was significantly decreased. By taking advantage of the double-quenching effect, this assay can detect PKA with a linear range of 0.05 to 5 U mL(-1) and a detection limit of 0.04 U mL(-1). In addition, the PKA inhibition assay and interferences experiments of CK2 and T4 PNK have been studied respectively. This assay was also successfully applied to PKA assay in serum samples and cell lysates, indicating that the developed method have the potential applications in protein kinase-related biochemical fundamental research and clinical diagnosis.

Amplified electrochemical detection of protein kinase activity based on gold nanoparticles/multi-walled carbon nanotubes nanohybrids.

A sensitive and simple electrochemical strategy has been developed for assay of protein kinase A (PKA) activity and inhibition using gold nanoparticles/multi-walled carbon nanotubes (AuNPs/MWNTs) nanohybrids. Key features of this assay included intrinsic peroxidase-like activity of positively-charged gold nanoparticles (+AuNPs) and signal transduction and amplification of multi-walled carbon nanotubes (MWNTs). In this assay, an N-terminally cysteine-containing peptide was self-assembled onto the gold electrode via Au-S bonding and used as substrate for PKA, and adenosine-5'-(γ-thio)-triphosphate was used as co-substrate. Upon thiophosphorylation in the presence of PKA, the AuNPs/MWNTs nanohybrids would be fixed onto the peptides via Au-S bond. The conjugated AuNPs/MWNTs nanohybrids could catalyze the 3, 3', 5, 5'-Tetramethylbenzidine (TMB) oxidation by H2O2 to form TMB oxidation product, which was reduced at the electrode surface to generate an electrochemical current. It was eT on state. The current signal intensity is proportional to the activity of PKA. Here, the presence of MWNTs not only increased the surface area for accumulation of +AuNPs but also could promote electron-transfer reaction. It was found that the electrochemical strategy can be employed to assay PKA activity with a low detection limit of 0.09 U/mL. The linear range of the assay for PKA enzymatic unit/ml was 0.1-1 U/mL. Furthermore, the interferences experiments of T4 polynucleotide kinase (T4 PNK) and Casein kinase II (CK2), and inhibition of PKA, have also been studied by using this strategy. The developed method would provide a diversified platform for kinase activity and inhibition monitoring.