双偏振干涉技术(DPI):生物分子相互作用研究的新工具

双偏振干涉(dual polarization interferometry,DPI)技术是近年来发展起来的一种免标记、实时、高灵敏和高分辨率的表面分析技术.它能够精确测量分子相互作用界面层的密度、厚度和质量的绝对值,可实时获取分子相互作用过程的动力学和结构信息.本文简单介绍了DPI的测量原理、仪器组成并对其与相关检测技术的对比进行了简要的概述;着重介绍了近10年来DPI技术在生物分子相互作用研究方面的应用进展,主要包括蛋白质之间以及与其他分子的相互作用,DNA与各种分子之间的相互作用,生物膜与其他分子的相互作用,蛋白质的吸脱附、聚集和结晶过程监测等;并对DPI技术未来的发展进行了展望.随着技术的不断发展,DPI将会在生物分析、纳米材料表征、能源相关表/界面研究等方面得到广泛应用.     

界面电场对血红蛋白在多孔金电极上的吸附和生物活性的影响(英文)

蛋白质的界面吸附及其生物活性因它在构建生物传感、生物电子器件和生物燃料电池等方面具有重要的作用而倍受关注.对此,界面电场是吸附的一个重要影响因素,它能明显地影响蛋白质分子在材料界面的吸附量、分子构象以及分子定向.本文应用电化学方法和红外光谱技术研究了血红蛋白在三维多孔金膜电极上的吸附动力学及其生物活性随界面电场的变化关系.结果表明,由界面电场产生的过量表面电荷可借助与蛋白质分子之间的静电作用加速蛋白质分子在电极表面的吸附,提高其吸附量;但是,过高的界面电场将破坏吸附蛋白质的构象以及降低它还原过氧化氢的催化活性;只有在零电荷电位下,吸附在电极表面的血红蛋白才能保持其天然的构象和生物催化活性.本研究将为生物传感器、生物电子器件和生物燃料电池的构建提供理论依据,加深对荷电生物界面上生物分子界面行为的认识.     

蛋白质膜界面流变行为研究进展

随着界面流变测量技术以及相关的光学辅助仪器的发展,近10年来界面流变学在食品、化妆品、医药等领域发挥了重要作用。本文介绍了近年来蛋白质膜界面流变行为的研究进展,着重介绍了蛋白质界面流变学在泡沫和乳液方面的研究。本文主要分为3个部分,内容包括:蛋白质膜界面流变学行为与泡沫、乳液稳定性的相互关系,蛋白质-多聚糖的界面流变行为研究,蛋白质-表面活性物质的界面流变行为研究。界面流变学在泡沫和乳液方面取得的较快研究进展不仅促进了人们对蛋白质膜界面流变行为更为深刻的理解,而且为更好地开发和应用蛋白质及其混合物作为表面活性物质和胶体稳定剂而提供重要的理论依据和指导。     

蛋白质界面取向的实验控制与表征

在生物材料、生物传感器、生物燃料电池等应用中都涉及到蛋白质界面吸附,而其中一个关键的科学问题就是蛋白质在基底界面上的吸附取向,控制好正确的取向是发挥蛋白质生物功能的关键。本文归纳和比较了各种用于控制蛋白质界面取向的固定化方法,总结了影响蛋白质界面吸附取向的因素,并对当前应用较多的表征蛋白质在界面吸附取向的方法进行了综述,最后指出蛋白质取向控制的机理研究将有助于推动相关应用领域的迅速发展。     

表面引发的DNA杂交链式反应的石英晶体微天平研究

通过石英晶体振荡技术研究了杂交链式反应这种核酸扩增的方法. 石英晶体微天平可以表征在晶体和溶液的界面上的DNA层, 并获得粘性穿透深度这一重要参数. 根据石英晶体表面吸附质量和振荡频率之间的关系, 我们测量了表面引发的杂交链式反应的动力学过程, 并获得界面上的粘度、剪切模量等参数. 这一工作为研究固液界面上核酸反应过程, 特别是杂交链式反应的机制提供了新的途径.     

可见-红外SFG方法及其在表面和界面表征研究中的应用

在近代表面科学的发展中,表面振动光谱学起着十分重要的作用.它在表征表面或界面上的被吸附物,研究被吸附物间和被吸附物与界面之间的相互作用等方面,是极为有效的方法和技术.     

蛋白功能化磁性纳米颗粒的制备及应用

蛋白功能化磁性纳米颗粒作为一种新型功能复合材料,已成为众多领域的研究热点。蛋白质在颗粒表面的稳定性、负载量及构象显著影响复合粒子的应用性能,而磁性纳米颗粒表面物化性质对颗粒的稳定性、分散性及磁性能对蛋白质的高效负载有重要影响。本文综述了磁性纳米颗粒表面修饰、蛋白功能化的方法以及蛋白在界面处构象变化的表征方法,介绍了蛋白功能化磁性纳米颗粒在酶催化合成、免疫分析检测及生物传感器等领域的应用,并对其未来的发展趋势进行了展望。     

基于蛋白质类淀粉样聚集的表面功能化

表面改性在各个领域都扮演着重要的角色,其在不改变材料本身性质的前提下,赋予材料新的性能和更高的价值.然而制约先进界面材料进一步应用发展的一个关键难题为缺少一种简单温和、高效环保和无色透明的普适性界面改性体系.自类淀粉样蛋白质用于界面改性被报道以来,该体系引起了学术界的广泛关注和研究,一系列不同形态结构的蛋白质基材料如纳米薄膜、纳米纤维、大颗粒聚集体(产物)、水凝胶及气凝胶等被成功发展.本综述首先阐述了蛋白质类淀粉样聚集的基本原理,然后总结了蛋白质类淀粉样聚集作为表面改性体系在生物医用涂层、分离/透析、生物矿化、柔性电子、智能织物、化学催化和环境污染物去除等方向的应用,最后指出该体系存在的不足并对未来发展方向进行了展望.     

扩张流变法研究表面活性剂在界面上的聚集行为

近年发展起来的界面流变测定技术在研究界面性质方面具有许多独特之处.本文结合我们的工作,总结了近年来有关该技术在表面活性剂界面聚集行为研究中的应用,讨论了扩张频率、表面活性剂浓度及疏水链长、无机盐和温度对表面扩张流变行为的影响,同时探讨了小分子表面活性剂与高分子表面活性剂表面扩张流变行为的区别以及小分子表面活性剂在气/液界面与液/液表面的扩张流变性的差异.大量研究表明,借助于界面流变性的测定不仅可以研究发生在界面上和界面附近的微观弛豫过程,而且可以探讨界面上超分子聚集体的形成,进而为乳状液和泡沫等分散体系的稳定性提供依据.     

BSA和SDBS在水溶液与空气界面的相互作用

表面活性剂与蛋白质在界面处的相互作用在许多应用里都会遇到．如去污、药品或生物制剂的制备和提纯、某些开科手术等等[1]这常常涉及到表面活性剂与蛋白质在界面的吸附，蛋白质大分子在界面吸附层的取向和构象．对于这两种物质在界面处可能发生的相互作用的性质，是表面活性剂     

Dual polarization interferometric and capillary electrophoretic analysis of supported lipid bilayer constructed on silica-based surface: Evaluation of its anti-protein adsorption effect

Supported lipid bilayer (SLB) has been demonstrated as a model of cell membranes with prospective bioanalytical or biotechnological applications. In this study, the formation of SLB and their potential biofunctionality against protein adsorption were investigated by Dual Polarization Interferometry (DPI) and Capillary Electrophoresis (CE). DPI studies on different formulations of double-chained, zwitterionic phospholipidlipids, allow the process of bilayer formation to be followed in situ and in real time. Furthermore the anti-protein adsorption effect provided by the various formulated SLBs was examined by DPI. In addition, the SLB coatings of the same lipid formulations were subsequently employed in CE experiments as a pseudo-stationary phase for demonstrating more efficient separation of alkaline protein standard mixtures. SLB-assisted CE was found to be capable of separating 4 alkaline proteins (protonated at neutral pH). This study demonstrates the applicability of DPI to monitor the process of SLB formation; and our findings, obtained by both DPI and CE, confirm that the presence of the SLB reduced drastically the problematic interactions between cationic, alkaline proteins and the negatively charged silica capillary wall, leading to better recovery and efficient separation of the proteins under investigation.     

A comparison between dual polarization interferometry (DPI) and surface plasmon resonance (SPR) for protein adsorption studies

This work was performed with the aim of comparing protein adsorption results obtained from the recently developed dual polarization interferometry (DPI) with the well-established surface plasmon resonance (SPR) technique. Both techniques use an evanescent field as the sensing element but completely different methods to calculate the adsorbed mass. As a test system we used adsorption of the lipase from Thermomyces lanuginosus (TLL) on C18 surfaces. The adsorbed amount calculated with both techniques is in good agreement, with both adsorption isotherms saturating at 1.30–1.35 mg/m² at TLL concentrations of 1000 nM and above. Therefore, this supports the use of both SPR and DPI as tools for studying protein adsorption, which is very important when comparing adsorption data obtained from the use different techniques. Due to the spot sensing in SPR, this technique is recommended for initial kinetic studies, whereas DPI is more accurate when the refractive index and thickness of the adsorbed layer is of more interest.     

Desolvation of BSA-ligand complexes measured using the quartz crystal microbalance and dual polarization interferometer

By taking advantage of their unique difference in hydration sensitivity, we have shown that dual polarization interferometer (DPI) and quartz-crystal microbalance with dissipation monitoring (QCM-D) measurements can be used together to explore the degree of desolvation involved in the binding of small drug molecules to an immobilized bovine serum albumin film in real time. Results with DPI and QCM-D show significantly different mass values for three ligands of varying hydrophobicities that may be attributed to changes in the degree of hydration of the ligand-protein complexes in accordance with the physicochemical properties of the ligands. Furthermore, our data suggest that masses measured by QCM-D can be overwhelmed by changes in water content of ligand-protein, binary complexes, which has important consequences for future studies using mechanical resonators to study protein-binding events.     

Organic functionalization of sidewall of carbon nanotubes

Using density functional theory, we have theoretically studied sidewall functionalization of carbon nanotubes (CNT) with a nucleophilic organic carbene, dipyridyl imidazolidene (DPI). When compared to the dissociated system, formation of the adduct from defect-free (5,5) tube and the DPI is weakly exothermic. However, introduction of (5,7,7,5) defect or nitrogen doping at the CNT stabilizes the adduct in both physical and chemical senses, suggesting a possible way to enrich the chemistry of sidewall functionalization. The work function of the adducts is found to decrease by approximately 0.4 eV per DPI/80 atoms. Upon binding of the DPI, electronic structures are modified in such a way that small gaps are introduced, where the size of the gap depends upon the degree of functionalization.     

Dual propagation inversion of truncated signals

Fourier transforms occur in a variety of chemical systems and processes. A few examples include obtaining spectral information from correlation functions, energy relaxation processes, spectral densities obtained from force autocorrelation functions, etc. In this article, a new functional transform, named the dual propagation inversion (DPI) is introduced. The DPI functional transform can be applied to a variety of problems in chemistry, such as Fourier transforms of time correlation functions, energy relaxation processes, rate theory, etc. The present illustrative application is to generating the frequency representation of a discrete, truncated time-domain signal. The DPI result is compared with the traditional Fourier transform applied to the same truncated time signal. For both noise-free and noise-corrupted time-truncated signals, the DPI spectrum is found to be more accurate, particularly as the signal is more severely truncated. In the DPI, the distributed-approximating-functional free propagator is used to propagate and denoise the signal simultaneously. Received: 30 January 2000 / Accepted: 6 July 2000 / Published online: 23 November 2000     

Photopolymerization of acrylamide initiated by the three‐component system safranine/triethanolamine/diphenyliodonium chloride: The effect of the aggregation of the salt

The effects of diphenyliodonium chloride (DPI) on the polymerization of acrylamide photoinitiated by the dye safranine with triethanolamine as a coinitiator were investigated in aqueous solutions. The salt notably increased the polymerization rate. This accelerating effects increased appreciably at concentrations of DPI at which the light scattering of the solutions became important. The effects of DPI on the photophysical properties of the dye were also investigated. Although the absorption and fluorescence were scarcely affected, the triplet yield increased by 60%. However, the polymerization rate increased by a factor much higher than that of the triplet yield. The results obtained at high concentrations of the salt could be ascribed to the presence of aggregates of the hydrophobic cations. The lower limit established for the formation of the aggregates was a DPI concentration of approximately 1 × 10⁻³ M. Possible mechanisms for the action of the salt were examined. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 4916–4920, 2004     

Application and mechanism of inhalation profile improvement of DPI formulations by mechanofusion with magnesium stearate

In our previous paper, we reported the inhalation properties of dry powder inhaler (DPI) formulations containing Compound A and mechanofusion-processed lactose carriers. The mechanofusion process with magnesium stearate (Mg-St) on the lactose carrier enhanced the fine particle fraction (FPF) value of the Andersen cascade impactor (ACI) study. The increase of FPF seemed to be associated with the increase of the dispersibility of drug particles. The objectives of this study were (1) to evaluate the applicability of lactose carrier mechanofusion-processed with Mg-St and (2) to examine the mechanism of FPF alteration by the mechanofusion process applied on the lactose carrier with or without additive. The inhalation profiles of DPI formulations containing four different pharmaceutical compounds were evaluated with an ACI. The dispersibility of the formulations was observed by particle size distribution measurement in the air stream and the adhesive force was measured bydirect separation method. It was found that higher FPF was obtained with lactose mechanofusion-processed with Mg-St as compared to control lactose carriers for all four compounds. This suggested that mechanofusion process with Mg-St is widely applicable in DPI formulations. The homogenization of surface adhesiveness was attributed to the increased FPF of the DPI including lactose mechanofusion-processed with Mg-St, as suggested by the combination of several physicochemical characteristics. Combination of different characterization methods would be of help to clarify the whole mechanism which defines the inhalation properties of DPI formulations.     

The salt-induced sphere-rod transition of micelles of dodecylpyridinium iodide in aqueous NaI solutions

Static light scattering has been measured on aqueous NaI solutions of dodecylpyridinium iodide (DPI) over NaI concentrations from 0 to 0.05 M. Reduced intensity of scattered light increases with increasing DPI concentration above the critical micelle concentration. The Debye plot is generally a curve with an initially positive slope and with a weakly convex, upward curvature, when the NaI concentration ranges from 0 to 0.005 M. The molecular weight of the spherical micelle of DPI is 28400 in water, and it increases slightly with increasing NaI concentration. The initial slope decreases with increasing NaI concentration and changes from positive to negative across 0.007 M NaI, which is the threshold for the sphere-rod transition and where the micelle has a molecular weight of 34400. At NaI concentrations from 0.01 to 0.05 M, the Debye plot is a curve with an initially negative slope and with a convex, downward curvature. The magnitudes of slope and curvature are larger, and the rodlike micelles of DPI have larger molecular weight and stronger mutual interaction, as the NaI concentration increases. The linear double logarithmic relationship between molecular weight and ionic strength holds for spherical and rodlike micelles, respectively.     

The role of diphenyl iodonium salt (DPI) in three‐component photoinitiator systems containing methylene blue (MB) and an electron donor

Three‐component initiators generally include a light‐absorbing photosensitizer, an electron donor that is often an amine, and the third component, which is usually an iodonium salt. To characterize the role of diphenyl iodonium chloride (DPI) in three‐component photoinitiator systems containing methylene blue (MB) as the photosensitizer, a systematic series of electron donors was used. The Rhem–Weller equation was used to verify the thermodynamic feasibility for photo‐induced electron transfer from the electron donors to the MB. Comparison of the photopolymerization rates of each two‐component initiator system (containing the photosensitizer and amine) to those of the corresponding three‐component system (with the addition of (DPI) allowed fundamental information regarding the role of the DPI to be obtained. It was concluded that the DPI enhances the photopolymerization kinetics in two ways: (1) it consumes an inactive MB neutral radical and produces an active phenyl radical, thereby regenerating the original methylene blue, and (2) it reduces the recombination reaction of the MB neutral radical and amine radical/cation. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 5863–5871, 2004     

Immobilization of enamel matrix derivate protein onto polypeptide multilayers. Comparative in situ measurements using ellipsometry, quartz crystal microbalance with dissipation, and dual-polarization interferometry

The buildup of biodegradable poly(L-glutamic acid) (PGA) and poly(L-lysine) (PLL) multilayers on silica and titanium surfaces and the immobilization of enamel matrix derivate (EMD) protein was followed by utilizing in situ ellipsometry, quartz crystal microbalance with dissipation, and dual-polarization interferometry (DPI). The use of the relatively new DPI technique validated earlier published ellipsometry measurements of the PLL-PGA polypeptide films. The hydrophobic aggregating EMD protein was successfully immobilized both on top of and within the multilayer structures at pH 5.0. DPI measurements further indicated that the immobilization of EMD is influenced by the flow pattern during adsorption. The formed polypeptide-EMD multilayer films are of interest since it is known that EMD is able to trigger cell response and induce biomineralization. The multilayer films thus have potential to be useful as bioactive and biodegradable coatings for future dental implants.