Enhanced Signal and Quantitative Detection of Anti-Interferon-Gamma Antibody by Using a Nanometer Biolinker.

For rapid screening and quantification of an antisera antibody, a nanometer bithiophene-based conductive biolinker can enhanced signal performance and can be used to verify the interaction of an anti-IFN-γ antibody with an IFN-γ protein. The experimental measurements take a generic approach which takes advantage of the functionality of thiophene-based linkers for biosensors. Effects associated with using bithiophene as a biolinker for surface plasmon resonance (SPR) spectroscopy are examined in this paper. By using an atomic force microscope (AFM), it was observed that the morphology of the bithiophene modified gold sensor surface became smoother than the original gold surface. We compared the response and concentration of the anti-IFN-γ antibody on a bithiophene-coated and dextran-coated biochip as well as on different thickness-modified surfaces under SPR relevant conditions. The results indicate that a response to IFN-γ molecules immobilized on a sensor using a bithiophene biolinker improved more than 8-fold when compared to that of a sensor using a dextran biolinker. Furthermore, the regeneration ability of the sensor surface shows good repeatability as only less than a 1% decrease was found after repeating the experimental work over 6 cycles. The characteristics provided us with a good platform for rapid screening, real-time monitoring and quantitative concentration of the autoimmune antibody activities.

Ligand-doped liquid crystal sensor system for detecting mercuric ion in aqueous solutions.

We developed a liquid crystal (LC) sensor system for detecting mercuric ion (Hg(2+)) in aqueous solutions. In this system, 4-cyano-4'-pentyl biphenyl (5CB) was doped with a sulfur- and nitrogen-containing ligand 5-(pyridine-4-yl)-2-(5-(pyridin-4-yl)thiophen-2-yl)thiazole (ZT) as the Hg(2+) specific LCs. When the system was immersed in the solution containing Hg(2+), the complex of ZT and Hg(2+) formed, which disrupted the orientation of LC and lead to a dark-to-bright transition of the image of LCs. From mercuric binding titrations monitored by UV-vis spectroscopy, it was found that 1:1 Hg(2+)/ZT complexes were formed. The limit of detection (LOD) of the system to Hg(2+) is 10 µM, and it did not respond to Cd(2+), Zn(2+), Cu(2+), Pb(2+), Fe(+), Mg(2+), Ca(2+), Na(+), and K(+). Besides, we also demonstrated that this system is capable of detecting Hg(2+) in tap water and pond water. Because the signal of this system is colorful under ambient light, which is readily understood by normal users, it can be used as a portable device to monitor the water quality at any location.

Development of a label-free impedance biosensor for detection of antibody-antigen interactions based on a novel conductive linker.

We developed a label-free impedance biosensor based on an innovative conductive linker for detecting antibody-antigen interactions. As the often used conventional long chain thiol is a poor conductor, it is not a suitable material for use in a faradaic biosensor. In this study, we adopted a thiophene-based conductive bio-linker to form a self-assembled monolayer and to immobilize the bio-molecules. We used cyclic voltammetry and impedance spectroscopy to verify the enhanced conductivity properties. Results showed that the electron transfer resistance of this new conductive linker was 3 orders of a magnitude lower than for a case using a conventional long chain thiol linker. With the decreased impedance (i.e. increased faradaic current), we can obtain a higher signal/noise ratio such that the detection limit is improved. Using fluorescence microscopy, we verified that our new conductive linker has a protein immobilization capability similar to a conventional long chain thiol linker. Also, using S100 proteins, we verified the protein interaction detection capability of our system. Our obtained results showed a linear dynamic range from 10 ng/ml to 10 µg/ml and a detection limit of 10 ng/ml. With our new conductive linker, an electrochemical impedance biosensor shows great potential to be used for point-of-care applications.

Significance of the pH-induced conformational changes in the structure of C-reactive protein measured by dual polarization interferometry.

Emerging evidence indicates that the conformation of C-reactive protein (CRP) plays important roles in human inflammation and cardiovascular disease (CVD). The different conformations in the structure of CRP under different pH conditions remain an important issue to be investigated for explaining various functions of CRP under certain physiologic and pathologic conditions. We directly measured the pH-induced conformational changes in the structure of CRP by dual polarization interferometry (DPI). The CRP was attached to an aldehyde-functionalized DPI sensor chip at a concentration of 50 µg/ml, and attained 2.019 ng/mm2 to form a surface coverage with a 1.71×10(-14) mol/mm2 CRP monolayer. A pentagonal structure with an average monolayer thickness value of 5.70±0.12nm and a layer density of 0.374±0.058 g/cm2 was obtained at pH 7.0. Moreover, the DPI biosensor signals directly reflected the considerable structural parameters and phenomena of conformational changes of CRP in a pH range of 2.0-10.0. The results obtained showed that the pentameric structure of CRP might dissociated into monomers or monomer aggregates as the pH shifts toward both acidic and alkaline conditions, but only partial rearrangements of CRP subunits might occur at extremely acidic physiological conditions. Considering the proinflammatory effect and subclinical chronic inflammation, pH-induced conformational changes in the structure of CRP between monomeric and pentameric formations may strongly relate to vascular atherosclerosis and subsequent CVD.

An efficient and highly selective deprotecting method for beta-(trimethylsilyl)ethoxymethyl ethers.

A series of beta-(trimethylsilyl)ethoxymethyl ethers were hydrolyzed to their corresponding alcohols in high yields by using a catalytic amount of CBr4 (15%) in MeOH under refluxing reaction conditions. The chemoselective deprotection between trialkylsilyl and beta-(trimethylsilyl)ethoxymethyl-protected alcohols can be achieved by using an alcohol with steric hindrance such as iPrOH. The selectivity also can be achieved in the CBr4/MeOH reaction mixture under ultrasonic reaction conditions.