Co-reporter:Shuai Zhao;Mo Yang;Wenfei Zhou;Baichao Zhang;Zhiqiang Cheng;Jiaxin Huang;Min Zhang;Zhiyou Wang;Rui Wang;Zhonglei Chen;Jinsong Zhu;Haitao Li
PNAS 2017 114 (35 ) pp:E7245-E7254
Publication Date(Web):2017-08-29
DOI:10.1073/pnas.1704155114
Chemical modifications on histones and DNA/RNA constitute a fundamental mechanism for epigenetic regulation. These modifications
often function as docking marks to recruit or stabilize cognate “reader” proteins. So far, a platform for quantitative and
high-throughput profiling of the epigenetic interactome is urgently needed but still lacking. Here, we report a 3D-carbene
chip-based surface plasmon resonance imaging (SPRi) technology for this purpose. The 3D-carbene chip is suitable for immobilizing
versatile biomolecules (e.g., peptides, antibody, DNA/RNA) and features low nonspecific binding, random yet function-retaining
immobilization, and robustness for reuses. We systematically profiled binding kinetics of 1,000 histone “reader–mark” pairs
on a single 3D-carbene chip and validated two recognition events by calorimetric and structural studies. Notably, a discovery
on H3K4me3 recognition by the DNA mismatch repair protein MSH6 in Capsella rubella suggests a mechanism of H3K4me3-mediated DNA damage repair in plant.
Co-reporter:Zhiqiang Cheng, Zhiyou Wang, Doreen E. Gillespie, Christopher Lausted, Zheng Zheng, Mo Yang, and Jinsong Zhu
Analytical Chemistry 2015 Volume 87(Issue 3) pp:1466
Publication Date(Web):January 12, 2015
DOI:10.1021/ac504110t
The application scope of surface plasmon resonance (SPR) and SPR imaging (SPRi) is rapidly growing, and tools such as high-performance and low-cost slides could enable more rapid growth of the field. We describe herein a novel silver slide, addressing the inherent instability of plain silver structure by improving adhesion between the glass substrate and the silver layer with a thin buffer layer of gold. Covered by a self-assembled monolayer (SAM) only, SPR characteristics of the slide remain steady for more than 3 months under regular storage. In a bioassay, the slide substantiates the predicted nearly 100% sensitivity improvement over gold slides and exhibits exceptional performance stability as determined by sensitivity and resolution measurements during the extended 40 000 s multicycle experiment. We demonstrate the suitability of this new slide for large-area SPRi, describing analysis results from a 1 296-ligand protein microarray. We predict this slide structure will provide a stable, high-sensitivity solution for high-throughput SPRi applications and other surface analysis platforms.
Co-reporter:Deependra Tyagi, Javier Batista Perez, Amita Nand, Cheng Zhiqiang, Peizhe Wang, Jie Na, Jingsong Zhu
Analytical Biochemistry 2015 Volume 471() pp:29-37
Publication Date(Web):15 February 2015
DOI:10.1016/j.ab.2014.11.001
Abstract
Surface plasmon resonance imaging (SPRi) has emerged as a versatile biosensor to detect a wide range of biomolecular interactions with divergent potential applications. However, the use of this advanced-level technology for stem cell lysate study is still not much explored. Cell lysates are significant biological analytes used for disease diagnostics and proteomic studies, but their complex nature limits their use as an analyte for SPRi biosensors. Here, we review the problems associated with the use of SPRi for stem cell lysate study and examine the role of surface chemistry, running buffer, and blocking solution in order to minimize nonspecific adsorption (NSA). We detect the expression of Oct4, Sox2, Nanog, Rex1, and Lin28 biomarkers present in mouse embryonic stem cell (mESC) lysate against their corresponding antibodies immobilized on the sensor surface with reduced NSA. The current study shows that the conjunction of SPRi and microarray can be used as a label-free, high-throughput, and rapid technique for detection of biomarkers and their relative abundance in stem cell lysate study.
Co-reporter:Javier Batista Perez, Deependra Tyagi, Mo Yang, Loany Calvo, Rolando Perez, Ernesto Moreno, Jinsong Zhu
Analytical Biochemistry 2015 Volume 484() pp:133-135
Publication Date(Web):1 September 2015
DOI:10.1016/j.ab.2015.05.005
Abstract
Self-assembled monolayers designed to immobilize capture antibodies are usually prepared using a mixture of functional and inactive linkers. Here, using low molar ratios (1:1 to 1:100) of the two linkers resulted in loss of binding capability of the anti-EGFR (epidermal growth factor receptor) antibody nimotuzumab, as assessed by surface plasmon resonance imaging. We then developed a simple theoretical model to predict the optimal surface density of the functional linker, taking into account the antibody size and linker diameter. A high (1:1000) dilution of the functional linker yielded the best results. As an advantage, this approach does not require chemical modification of the protein.
Co-reporter:Zhiyou Wang, Zhiqiang Cheng, Vikramjeet Singh, Zheng Zheng, Yanmei Wang, Shaopeng Li, Lusheng Song, and Jinsong Zhu
Analytical Chemistry 2014 Volume 86(Issue 3) pp:1430
Publication Date(Web):December 29, 2013
DOI:10.1021/ac402126k
The silver surface plasmon resonance (SPR) sensor has long been explored due to its intrinsic sensitivity enhancement over the conventional single-layered gold SPR sensor. However, the silver SPR sensor has not been exploited for practical applications because of pronounced instability problems. We propose a novel gold–silver–gold trilayered SPR sensor chip, in which an extra buffer layer of gold is added between the silver and substrate adhesion layer (i.e., chromium) compared to the previously reported silver–gold bilayered SPR sensors. Subjected to prolonged agitation in phosphate-buffered saline (PBS) solution, the new chip exhibited high integrity according to both optical and atomic force microscopy (AFM) analysis. Having undergone repeated cycles of calibration, binding, and regeneration in various chemical solutions, 25 regions of interest (ROIs) over a 14 mm ×14 mm area were chosen and monitored by large detection area SPR microscopy; the new sensor chip exhibited stability comparable to the single gold layered SPR chip. In terms of sensing performances, over 50% increases in sensitivity and signal-to-noise ratio (S/N) than those of the single gold layered SPR chip were determined by SPR microscopy at 660 nm. Protein arrays of protein A and bovine serum albumin (BSA) on both the new chip and single-layered gold SPR chip were fabricated and underwent biomolecular interactions with human IgG, for the purpose of consistency, comparison on kinetics parameters, values from the microarray trilayered chip showed reasonable consistency with those from the single gold layered SPR chip. This study suggests that the new chip is a viable alternative to the conventional single gold layered SPR chip with improved sensing performances.
Co-reporter:Amita Nand, Vikramjeet Singh, Javier Batista Pérez, Deependra Tyagi, Zhiqiang Cheng, Jingsong Zhu
Analytical Biochemistry 2014 Volume 464() pp:30-35
Publication Date(Web):1 November 2014
DOI:10.1016/j.ab.2014.06.002
Abstract
In recent years, in situ protein synthesis microarray technologies have enabled protein microarrays to be created on demand just before they are needed. In this paper, we utilized the TUS-TER immobilization technology to allow label-free detection with real-time kinetics of protein–protein interactions using surface plasmon resonance imaging (SPRi). We constructed an expression-ready plasmid DNA with a C-terminal TUS fusion tag to directionally immobilize the in situ synthesized recombinant proteins onto the surface of the biosensor. The expression plasmid was immobilized on the polyethylene imine-modified gold surface, which was then coupled with a cell-free expression system on the flow cell of the SPRi instrument. The expressed TUS fusion proteins bind on the surface via the immobilized TER DNA sequence with high affinity (∼3–7 × 10−13 M). The expression and immobilization of the recombinant in situ expressed proteins were confirmed by probing with specific antibodies. The present study shows a new low cost method for in situ protein expression microarrays that has the potential to study the kinetics of protein–protein interactions. These protein microarrays can be created on demand without the problems of stability associated with protein arrays used in the drug discovery and biomarker discovery fields.
Co-reporter:Amita Nand, Vikramjeet Singh, Peizhe Wang, Jie Na, Jingsong Zhu
Analytical Biochemistry 2014 Volume 465() pp:114-120
Publication Date(Web):15 November 2014
DOI:10.1016/j.ab.2014.07.028
Abstract
Lectin microarrays have emerged as a novel platform for glycan analysis during recent years. Here, we have combined surface plasmon resonance imaging (SPRi) with the lectin microarray for rapid and label-free profiling of stem cells. In this direction, 40 lectins from seven different glyco-binding motifs and three different cell lines—mouse embryonic stem cells (mESCs), mouse-induced pluripotent stem cells (miPSCs), and mouse embryonic fibroblast stem cells (MEFs)—were used. Pluripotent mouse stem cells were clearly distinguished from non-pluripotent stem cells. Eight lectins—DBA, MAL, PHA_E, PHA_L, EEL, AAL, PNA, and SNA—generated maximal value to define pluripotency of mouse stem cells in our experiments. The discriminant function based on lectin reactivities was highly accurate for the determination of stem cell pluripotency. These results suggested that glycomic analysis of stem cells leads to a novel comprehensive approach for quality control in cell-based therapy and regenerative medicine.
Co-reporter:Tianyi Zhao, Dongmei Zhang, Chunmei Ding, Jinglun Zhou, Jinsong Zhu, Lei Jiang
Particuology 2014 Volume 17() pp:11-14
Publication Date(Web):December 2014
DOI:10.1016/j.partic.2014.04.006
•A multi-functional polycarbonate film was prepared by a simple casting process.•The polymer film showed highly hydrophobicity, heat-resistance and transparency.•The high hydrophobicity could be retained up to 390 °C.•The polymer film may have potential applications in aerospace and food industries.A multi-functional polymer film with high hydrophobicity and transparency was formed by simple casting of a polycarbonate solution onto a substrate having micro-scale roughness. The high hydrophobicity was heat-resistant, which can be retained up to 390 °C. The polymer film may have potentially wide-ranging applications in industry and high technology.
Co-reporter:Kun Wang, Yuhang Wan, Zheng Zheng, Fuchu He, Zhiyuan Hu, Jinsong Zhu
Sensors and Actuators B: Chemical 2012 Volume 169() pp:393-396
Publication Date(Web):5 July 2012
DOI:10.1016/j.snb.2011.12.030
Voltage scanning surface plasmon resonance (VSSPR) detection is realized by a metal/electro-optic dielectric/metal (MEM) three-layer sensor chip used in a prism-coupled attenuated total reflection (ATR) setup. Sweeping DC voltage is applied and recorded in real time with a fixed incident angle near the resonant angle. The VSSPR curve is defined by the reflected intensity and the applied voltage. It is observed experimentally that the resonant voltage obtained by the centroid method varied linearly with the concentration of the analytes and can be used as a characteristic and quantitative parameter to describe the SPR effect. This method can be used for refractive index detection, and also have potential applications in surface plasmon resonance imaging systems.
Co-reporter:Xin Ma, Xinlong Xu, Zheng Zheng, Kun Wang, Yalin Su, Jiangfeng Fan, Rui Zhang, Lusheng Song, Zhiyou Wang, Jinsong Zhu
Sensors and Actuators A: Physical 2010 Volume 157(Issue 1) pp:9-14
Publication Date(Web):January 2010
DOI:10.1016/j.sna.2009.11.004
An electro-optically modulated intensity interrogation method based on tunable waveguide coupled surface plasmon resonance sensors has been proposed. It has been theoretically and experimentally demonstrated that the proposed scheme can enable sensitive measurement of measurand variations. By modulating the refractive index in the waveguide layer, this interrogation method yields modulated signal whose amplitude is related to measurand's refractive index. This amplitude modulated signal offers a higher signal to noise ratio and eliminates additive noise in the sensor system. A preliminary investigation using saline buffers with different NaCl concentrations shows a resolution of 2.3 × 10−6 refractive index unit by our approach. Resolution can be controlled by the amplitude of the applied modulation voltage and can be further enhanced by optimizing the device structure or improving the electro-optical (E-O) coefficient of the E-O material. This approach is simple, stable, and promising for low-cost or multi-channel SPR biosensor applications.