DETERMINATION OF BOVINE SERUM ALBUMIN CONJUGATED DRUGS BY IMMOBILIZATION AS MICROARRAYS AND OBLIQUE-INCIDENCE REFLECTIVITY DIFFERENCE MICROSCOPY

Human serum is a mixture of various proteins which may interact with drugs. Therefore, it is of interest to investigate the binding kinetics of pharmaceuticals with their corresponding antibodies in serum. In this article, microarrays and a label-free biosensor were used to study these interactions. Microarrays provide a high-throughput platform for characterizing biomolecular interactions, and the label-free oblique-incidence reflectivity difference biosensor avoids the drawbacks of fluorescence-based methods. The experimental results show that the binding affinities between most of the drugs and their antibodies were reduced in human serum because the bulky proteins block the access to or reduce the stability of the reaction complexes. Therefore, one should be mindful when in vitro or in vivo testing the efficiency of potential drugs and their antibodies.     

Evaluation of kinetics using label-free optical biosensors

Optical biosensors provide a platform for qualitatively and quantitatively analyzing various biomolecular interactions. In addition to advantages such as label-free and high-throughput detection, these devices are also capable of measuring real-time binding curves in response to changes in optical properties of biomolecules. These kinetic data may be fitted to models to extract binding affinities such as association rates, dissociation rates, and equilibrium dissociation constants. In these biosensors, one of the binding pair is usually immobilized on a solid substrate for capturing the other. Due to the nature of these surface-based methods, mass transport effects and immobilization heterogeneity may cause problems when fitting the kinetic curves with the simple one-to-one Langmuir model. Here, real-time binding curves of various antibody–antigen reactions were obtained using an ellipsometry-based biosensor, and the results were fitted to the simple one-to-one model as well as a more sophisticated approach. The results show that the one-to-two model fitted much better to the curves than the one-to-one model. The two-site model may be explained by assuming two immobilization configurations on the surface. In summary, in fitting real-time curves obtained from optical biosensors, more sophisticated models are usually required to take surface-related issues, such as immobilization heterogeneity, and mass transport effects within targets, into account.     

Real-time,label-free characterization of oligosaccharide-binding proteins using carbohydrate microarrays and an ellipsometry-based biosensor

Carbohydrates present on cell surfaces mediate cell behavior through interactions with other biomolecules. Due to their structural complexity, diversity, and heterogeneity, it is difficult to fully characterize a variety of carbohydrates and their binding partners. As a result, novel technologies for glycomics applications have been developed, including carbohydrate microarrays and label-free detection methods. In this paper, we report using the combination of oligosaccharide microarrays and the label-free oblique-incidence reflectivity difference microscopy for real-time characterization of oligosaccharide-binding proteins. Aminated human milk oligosaccharides were immobilized on epoxy-coated glass substrates as microarrays for reactions with Family 1 of solute-binding proteins from Bifidobacterium longum subsp. infantis (B. infantis). Binding affinities of these protein–oligosaccharide interactions showed preferences of Family 1 of solute-binding proteins to host glycans, which helps in characterizing the complex process of human milk oligosaccharides foraging by B. infantis.     

DETECTION OF FORMATION AND DISINTEGRATION OF MICELLES BY OBLIQUE-INCIDENCE REFLECTIVITY DIFFERENCE MICROSCOPY

An oblique-incidence reflectivity difference (OI-RD) scanning microscope was developed for label-free detection of the formation and disintegration of micelles upon solid substrates. Micelles are made of polymers with hydrophilic heads in contact with the surrounding solvent and hydrophobic tails in the micelle center. This characteristic makes them very efficient drug carriers. Streptavidin molecules were first printed on glass slides for capturing biotinylated polymers. Micelles were formed when the concentration of polymers was higher than a critical value. The formation of micelles resulted in an increase in the oblique-incidence reflectivity difference signals. As the concentration of polymers decreased below the critical value, micelles were disintegrated, and a corresponding decrease in the oblique-incidence reflectivity difference signals was observed. This microscope was employed for the real-time monitoring of the formation and disintegration of two different micelles. The critical concentration above which micelles were formed was determined to be around 0.0006 mg/mL for micelles made of PEG^5KCA₈ polymers. The results suggest that this microscope would have practical application in testing the efficiency and durability of micellar drug carriers.     

A MINIATURE SURFACE PLASMON RESONANCE BIOANALYTICAL SYSTEM FOR FIELD DETECTION OF MICROCYSTIN-LR IN SURFACE WATER

A miniature surface plasmon resonance (SPR)–based immunoassay system for analyzing microcystin-LR (MC-LR) in surface water has been developed. A Spreeta biosensor was adopted in the SPR flow injection analysis (FIA) system. An optional biofilm immobilization method was exploited using covalent-coupling between a bioconjugate of microcystin-LR and bovine serum albumin [(MC-LR)-BSA] and the golden surface. An acceptable relative standard deviation of repeatability, 1.29% (n = 20), was achieved. An analysis of the calibration curve showed that this method has a detection limit of 0.55 ng/mL, a half-maximal inhibitory concentration (IC₅₀) of 5.6 ng/mL, and a quantitative range of 1.56–25.00 ng/mL. The recovery from water samples containing varying concentrations of MC-LR ranged from 90% to 112%. After samples of tap water and pool water were assayed, it was concluded that tap water did not contain a measurable amount of the analyte, whereas pool water contained 2.39 ng/mL. Compared to other methods of analysis, SPR immunoassay has the advantage of being label-free, rapid, and cost-effective and can easily detect levels of MC-LR within the World Health Organization (WHO) drinking water guidelines. The SPR FIA system with reusable biosensor is compact and convenient for instant in situ determination of MC-LR levels in environmental water.     

AN OBLIQUE-INCIDENCE REFLECTIVITY DIFFERENCE STUDY OF THE DEPENDENCE OF PROBE-TARGET REACTION CONSTANTS ON SURFACE TARGET DENSITY USING STREPTAVIDIN-BIOTIN REACTIONS AS A MODEL

A combination of the microarray platform and oblique-incidence reflectivity difference (OI-RD) microscopy was used to study the dependence of kinetic constants of probe-target reactions on the surface density of immobilized targets. Streptavidin-biotin reactions with a very high binding affinity were employed as the study model. Oblique-incidence reflectivity difference microscopy, a label-free and surface-based detection technique, was developed for monitoring real-time binding curves between two interactive biomolecules, enabling the acquisition of kinetic constants such as on-rate, off-rate, and equilibrium dissociation constants. These kinetic constants are important in characterizing biomolecular interactions because in living cells all intercellular and intermolecular reactions are at dynamic rather than at stable equilibrium. The kinetic constant of streptavidin binding to surface-immobilized biotin-bovine serum albumin was demonstrated to be significantly affected by the density of surface bovine serum albumin conjugates, mainly due to mass-transport effects within targets.     

表面等离子体共振理论仿真研究

为了掌握影响激发表面等离子体共振(SPR)现象的因素,文中以薄膜光学理论为基础,利用Winspall软件模拟研究Kretschman结构下,金膜厚度、待测介质折射率以及三棱镜类型对表面等离子体共振吸收峰位变化的影响,得出了最佳膜厚以及影响激发SPR现象的棱镜折射率和待测物质折射率随共振角度变化的规律,为SPR传感器的设...     

REVIEW: ADVANCES AND APPLICATIONS OF SURFACE PLASMON RESONANCE BIOSENSING INSTRUMENTATION

Surface plasmon resonance biosensor technology has enjoyed rapid development in the past two decades and undergone a boom in recent years. The growing demand for analytical applications has brought a variety of these instruments on to the market, and these sensors are nearly essential in the biochemical analysis laboratory, with wide applications in the life sciences, environmental monitoring, food safety, drug screening, and clinical diagnosis. After examining the last three years of literature on surface plasmon resonance biosensor instruments, in this article excellent studies are reviewed that have practical significance for progress in the instrumentation. Among the studies on improving sensor performance, the focus has been on metal thin film modification and data processing; regarding innovative design, the combinations of biosensing techniques with microfluidic chips and electrochemical with surface-enhanced Raman scattering are discussed. Instrument miniaturization has also been considered, and some personal perspectives are presented. In addition, various applications of these sensors are in biochemistry, medical diagnostics, food safety, and environmental monitoring. Finally, conclusions on the developments to date are summarized and future development trends are proposed.     

基于表面等离子体共振技术油水持率传感器的实验研究

为研究表面等离子体共振技术在油井产液剖面持率参数测试方面的潜在应用,采用磁控溅射方法在三棱镜底面沉积不同厚度的金膜,制作Kretschman型表面等离子共振传感元件,对不同体积浓度的油水折射率进行测定。实验结果表明,对于折射率相同的三棱镜,沉积的金膜厚度越大,共振吸收峰越弱。表面等离子体共振峰位对不同体积浓度的油水介质很敏感,介质体积浓度由低到高变大时,表面等离子体共振吸收峰向右平移,共振角逐渐增大。实验表明,表面等离子体共振传感测试技术对不同浓度油水介质的折射率变化非常敏感,能够用其进行流体持率的测量。     

液体折射率测定方法分析

折射率是反映介质光学性质的一个重要参数,对其精确测定有重要的意义。文章对液体折射率测量方法做了归纳和分类,并对几种主要测定方法做了介绍,分析了各自的特点和存在的问题。尤其对基于表面等离子体波共振效应的新型光纤传感系统的结构、工作原理及其在液体折射率测定等方面应用做了具体的分析和介绍。     

镀金属膜长周期光纤光栅的传感特性

本文基于耦合模理论, 建立了严格的四层金属膜理论模型, 探讨了镀金属膜长周期光纤光栅(Long-period fiber grating, LPFG)的温度、 应变及折射率特性, 以及镀膜参数对镀金属膜长周期光纤光栅光谱特性的影响.仿真结果表明, 长周期光纤光栅表面最优的金属膜厚度将引起表现等离子共振(Surface-plasmon resonance, SPR)特性, 这一特性将使得LPFG 对稳定及折射率都有较高的敏感性, 而对应变影响较小. 理论分析表明, 银膜厚度在0.8-1.2 nm 范围内时, 折射率敏感度达到最大值为42.402 6, 敏感度增加4.5%. 仿真结果为镀膜长周期光纤光栅的设计及参数优化提供了理论指导.     

双金属核壳纳米结构中荧光单分子的表面能量转移及金属操控自发辐射效应

贵金属表面附近的荧光分子因其表面等离子体共振的影响,荧光发射特性发生显著变化,被广泛应用在荧光探针等纳米器件的设计、开发中。荧光分子与金属之间的能量转移机制是设计此类荧光探针的基础。使用时域有限差分(FDTD)方法,对Au/SiO₂/Ag核壳纳米复合结构的等离子体杂化场中荧光单分子的表面能量转移(SET)效应和金属操控自发辐射效应进行了理论仿真研究。研究了金核和银壳共同作用时,荧光分子的SET和金属调控自发辐射过程随荧光分子位置及分子偶极矩取向的变化规律。计算结果表明,由于金核和银壳之间的局域表面等离子体共振杂化耦合,荧光分子与金属间的能量转移效率与距离d呈现出10次方的关系,这一结果明显区别于常规的荧光共振能量转移(FRET)效应,较之单金属结构的SET效应更加剧烈。这一结果有希望在生物光子学领域的纳米级局域光源的创建和生物分子的检测中得到应用。     

角度指示型表面等离子共振传感器的研究

表面等离子体共振(SPR)技术被广泛应用于生物化学传感、药物分析和环境监测等领域。现基于表面等离子共振原理,在固定入射光波长为632.8nm的He-Ne激光入射下,采用镀有50nm银膜的Kretschmann棱镜型结构作为耦合器件,实现角度指示型SPR传感器对纯净水和酒精水溶液的检测。实验结果发现该SPR传感器具有较好的稳定性,其角度灵敏度达到219°/RIU。     

The fabrication of functional biosurface composed of iron storage protein, ferritin

A functional biosurface applicable to a biomemory device was fabricated using ferritin, which is one of the globular protein complexes consisting of 24 protein subunits, which can be classified as metalloproteins. For the fabrication of uniform ferritin layer, 11-MUA(11-mercaptoundecanoic acid) was used as a linker material. The formation of the ferritin layer was confirmed by surface plasmon resonance (SPR) spectroscopy, and the morphology of the immobilized ferritin was analyzed by scanning tunneling microscopy (STM). The electrochemical redox property investigation was accomplished by the cyclic voltammetry (CV) technique. These results of adsorbed ferritin on the modified electrode can be used for the fabrication of bioelectronics.     

表面等离子体谐振（SPR）生化分析仪的研制

表面等离谐振(Surface Plasmon Resonance,SPR)生化分析仪是一种基于物理光学原理的新型生化分析系统,SPR技术作为一种表在反应的生物传感技术,具有很重要的生化分析能力,可用于无标记实时监测许多种类生物分子之间的反应.SPR生物分析技术已成为直接实时观测生物分子间相互作用的主导技术.近年来SPR传感技术受到重点研究和发展;SPR生化分析仪也已经商品化.在本文中详细介绍了自行研制的新型SPR-2000型生化分析仪,仪器的主要性能是入射角扫描范围40°～70°,测角精度优于1×10-3度,探测折射率范围1.04～1.47,温度范围4℃～60℃、检测灵敏度可达10-11M等.利用该系统实时测试了1gG-GAHIgG的动态反应过程.实验证明该分析仪精度和灵敏度高,适用范围宽,并已提供多家使用,具有比较好的商品化性能.中国图分类号 TH744     

一种降低坐标测量机定位误差的方法

提出了一种将实际测量数据点与工件的CAD模型理论数据点优化匹配的算法。使用该算法可有效提高工件设计坐标系相对于坐标测量机测量坐标系的定位精度,可广泛用于精密测量时测量结果的比较分析。     

EFFECT OF MICROWAVE TREATMENT ON WC INSERTS FOR DRILLING OF GFRP COMPOSITES

Due to excellent properties such as high specific strength, high specific modulus, and corrosion resistance, Glass Fiber Reinforced Plastics (GFRP) find wide applications particularly in the aerospace and automotive industries. In order to fabricate structural components of GFRP to near-nett shape, machining of GFRP composites is necessary. Drilling is the most common machining operation for GFRP composites. The fiber pull-out, cracking of matrix during the entry and exit of drill, rapid tool wear, and hole shrinkage are problems associated with machining of GFRP. The present study attempts an innovative idea of improving the cutting performance of Tungsten carbide (WC) inserts (K-20) by post-sintering using microwave irradiation.     

GGBP蛋白修饰的表面等离子共振微创血糖检测仪

根据基于微流控芯片的组织液透皮抽取系统设计了一种小型化微创人体血糖检测仪器.该仪器基于微创的方法,利用真空负压抽取人体组织液,并采用表面等离子共振(SPR)技术,通过检测皮肤真皮层组织液中的葡萄糖浓度来预测血液中的葡萄糖浓度.通过绑定对葡萄糖具有特异性吸附的D-半乳糖/D-葡萄糖结合蛋白(D-GGBP),对SPR传感器表面进行预处理,实现对葡萄糖分子的特异性吸附.实验配制了不同浓度的葡萄糖溶液,检测并得出葡萄糖溶液浓度与折射率的关系曲线.应用课题组设计的微创血糖检测仪,实验测量了葡萄糖溶液浓度与组织液浓度,并与血糖仪测量得到的葡萄糖溶液浓度进行了比较.结果表明,使用GGBP修饰过的表面等离子共振传感器测量葡萄糖水溶液浓度的下限为0.625 mg/dL,当葡萄糖水溶液浓度在0.625～5 mg/dL时有较好的线性.通过测试实验验证了该仪器的可行性,显示了结合GGBP蛋白的SPR测量技术在微创血糖检测领域有良好应用前景.     

应力脉冲对CCCD-SHPB试验系统测试结果影响的数值分析

3维动态有限元分析结果表明,脉冲上升沿时间、幅值及试件直径对CCCD-SHPB(central cracked circular disk-split Hopkinson pressure bar)动态断裂试验系统的试验结果有较大影响,改变脉冲上升沿时间或脉冲幅值可以得到不同加载速率下材料的动态断裂韧度.进一步计算结果表明,在动态加载的初期阶段,由于试件两边载荷不平衡,导致CCCD-SHPB动态断裂试验系统中存在与试件尺寸相关的特征时间ΔT,且ΔT近似等于应力波在试件中来回往返两次的时间;动态断裂试验时,试件从加载至断裂的时间应大于特征时间ΔT,否则试验结果不可靠.     

OPTIMAL MQL AND CUTTING CONDITIONS DETERMINATION FOR DESIRED SURFACE ROUGHNESS IN TURNING OF BRASS USING GENETIC ALGORITHMS

The evolving concept of minimum quantity of lubrication (MQL) in machining is considered as one of the solutions to reduce the amount of lubricant to address the environmental, economical and ecological issues. This paper investigates the influence of cutting speed, feed rate and different amount of MQL on machining performance during turning of brass using K10 cemented carbide tool. The experiments have been planned as per Taguchi's orthogonal array and the second order surface roughness model in terms of machining parameters was developed using response surface methodology (RSM). The parametric analysis has been carried out to analyze the interaction effects of process parameters on surface roughness. The optimization is then carried out with genetic algorithms (GA) using surface roughness model for the selection of optimal MQL and cutting conditions. The GA program gives the minimum values of surface roughness and the corresponding optimal machining parameters.