氨基酸在固体表面的吸附*

介绍了氨基酸在固体表面吸附的常见吸附模式和吸附等温线类型。在常见的等温式中Sips等温式能处理液相中吸附质浓度从低到高、吸附剂表面从均匀到不均匀等各种情形下的固/液吸附体系,得到的等温线一般归属4类：S型、L型、H型和C型;总结和讨论了各吸附参数对氨基酸吸附的影响,具体考察了氨基酸种类、固体表面的性质、溶剂种类、介质的pH、介质离子强度和温度等因素对氨基酸吸附的影响,指出通过优化各吸附参数,可以调整氨基酸和固体吸附剂之间的静电吸引力或疏水作用,达到最好的吸附效果;对氨基酸的固体吸附剂进行了分类阐述。     

表面等离子体共振技术在电化学反应过程研究中的应用

简述了表面等离子体共振(SPR)的基本原理,并综述了表面等离子体共振技术在电化学反应过程中的应用。SPR技术可以无需任何标记原位实时地检测分子间的相互作用,也可用于连续监测吸附/脱附和缔合/解离过程。表面等离子共振光谱(SPRS)与电化学技术结合可用来同时表征和处理电极/溶液的界面,在电化学掺杂/去掺杂过程、吸附/脱附反应的研究、痕量物质的检测、薄膜厚度、介电常数的测定等方面的应用已取得了很大的进展。     

蛋白质在固体表面吸附的研究进展

蛋白质在固体表面的吸附有多种理论模型和实验分析.蛋白质吸附主要包括分子传递、吸附、重排、交换、解吸等步骤.蛋白质在表面的状态由表面性能、静电作用及蛋白质自身性质等因素决定.蛋白质分子在界面吸附后发生构象改变,引起熵增.     

贵金属纳米材料的液相合成及其表面等离子体共振性质应用

金属中的表面等离子体共振是描述其导带电子在电磁场作用下集体振荡的一个物理概念。金属纳米颗粒由于其表面等离子体共振性质表现出独特的光学应用。本文在相关研究的基础上,综述了具备表面等离子体共振性质的不同形貌及多种复合结构的贵金属纳米材料的制备和应用,并对其发展及应用前景进行了展望。     

贵金属纳米材料的液相合成及其表面等离子体共振性质应用

金属中的表面等离子体共振是描述其导带电子在电磁场作用下集体振荡的一个物理概念。金属纳米颗粒由于其表面等离子体共振性质表现出独特的光学应用。本文在相关研究的基础上,综述了具备表面等离子体共振性质的不同形貌及多种复合结构的贵金属纳米材料的制备和应用,并对其发展及应用前景进行了展望。     

金纳米棒对表面等离子体耦合发射的增强作用研究

局域型与传播型表面等离子体相互作用引起的电磁场增强是调控表面等离子体耦合发射(SPCE)的有效方法。通过静电吸附将金纳米棒(AuNRs)引入光滑金基底表面,再匀胶625 nm发射的量子点荧光薄膜,检测到高度p偏振的定向辐射;借助体系独特的等离子体耦合特性,相较于常规SPCE和自由空间发射分别实现了约40和150倍的信号增强。研究结果表明,信号调控主要来自AuNRs的局域等离子体共振、体系的增强等离子体电磁场以及“热点”结构的形成对处在耦合猝灭区的荧光团带来的“去猝灭”效应。通过条件优化,探究了AuNRs调控SPCE信号的影响因素,包括AuNRs的修饰浓度以及量子点荧光薄膜的厚度,为高效耦合增强体系的构建提供了研究方法。等离子体与不同波长的荧光发射耦合作用各异,将AuNRs用于多波长SPCE体系的信号调控,在不同的发射角度实现了对应波长的耦合增强检测。AuNRs修饰过程简单,增强效果显著,是一种调控等离子体耦合发光的普适方法,有望提高体系的检测性能;结合SPCE的波长分辨特性,可为构建多波长同时增强检测体系提供可行的光谱分析手段。     

苯基取代乙烯类化合物在硅胶表面敏化光氧化反应的研究

随着人们对吸附态有机分子光谱性质的研究的不断深入,光化学中的一个重要方面——表面光化学,也得到了很大的发展。在固体表面上,人们观察到了许多与溶液中不同的现象。这主要是由于固体表面上常常表现出某种特有的吸附特性,从而对被吸附物质的能级状态和分子结构产生影响,进而改变了它们的反应性能,这类报道已有很多^[1]。     

贵金属纳米材料的液相合成及其表面等离子体共振性质应用

金属中的表面等离子体共振是描述其导带电子在电磁场作用下集体振荡的一个物理概念.金属纳米颗粒由于其表面等离子体共振性质表现出独特的光学应用.本文在相关研究的基础上,综述了具备表面等离子体共振性质的不同形貌及多种复合结构的贵金属纳米材料的制备和应用,并对其发展及应用前景进行了展望.     

表面等离子体共振技术在蛋白质-蛋白质相互作用研究中的应用

表面等离子共振(SPR)近年来迅速发展为用于分析生物分子相互作用的一项技术.该技术无需标记、特异性强、灵敏度高、样品用量小,可实现在线连续实时检测.目前SPR已被广泛应用于免疫学、蛋白质组学、药物筛选、细胞信号转导、受体/配体垂钓等领域.该文阐述了基于表面等离子体共振技术生物传感器的基本原理和技术流程,综述了SPR在蛋白质-蛋白质相互作用动力学研究、蛋白质结构及功能研究、蛋白质突变和碎片分析、信号转导中的应用以及SPR在蛋白质-蛋白质相互作用研究中的多项关键技术.指出SPR通过与光谱、电化学等多技术联用后,可以获得更加详实的信息.     

氢原子在Ti(0001)表面吸附的密度泛函理论研究

用密度泛函理论研究了氢原子的污染对于Ti(0001)表面结构的影响. 通过PAW总能计算研究了p(1×1)、p(1×2)、3^1/2×3^1/2R30[deg]和p(2×2)等几种氢原子覆盖度下的吸附结构, 以及在上述结构下Ti(0001)面fcc格点和hcp格点的氢原子吸附. 结果表明, 在p(1×1)-H、p(1×2)-H、3^1/2×3^1/2R30[deg]-H和p(2×2)-H几种H原子覆盖度下, 以p(1×1)-H结构的单个氢原子吸附能为最大. 在p(1×1)-H吸附结构下, 由于氢原子吸附导致的Ti(0001)表面Ti原子层收缩的理论计算数值分别为-2.85%(hcp吸附)和-4.31%(fcc吸附), 因此实际上最有可能的情况是两种吸附方式都有一定的几率. 而实验中观察到的所谓“清洁”Ti(0001)表面实际上是有少量氢原子污染的表面. 不同覆盖度和氢分压下, 氢原子吸附的污染对Ti(0001)表面结构有极大的影响, 其表面的各种特性都会随覆盖度的不同而产生相应的变化.     

Monte Carlo simulations of antibody adsorption and orientation on charged surfaces

Monte Carlo simulations were performed to study the adsorption and orientation of antibodies on charged surfaces based on both colloidal and all-atom models. The colloidal model antibody consists of 12 connected beads representing the 12 domains of an antibody molecule. The structure of the all-atom antibody model was taken from the protein databank. The effects of the surface charge sign and density, the solution pH and ionic strength on the adsorption and orientation of different colloidal model antibodies with different dipole moments were examined. Simulation results show that both the 12-bead and the all-atom models of the antibody, for which the dipole moment points from the Fc to (Fab)2 fragments, tend to have the desired "end-on" orientation on positively charged surfaces and undesired "head-on" orientation on negatively charged surfaces at high surface charge density and low solution ionic strength where electrostatic interactions dominate. At low surface charge density and high solution ionic strength where van der Waals interactions dominate, 12-bead model antibodies tend to have "lying-flat" orientation on surfaces. The orientation of adsorbed antibodies results from the compromise between electrostatic and van der Waals interactions. The dipole moment of an antibody is an important factor for antibody orientation on charged surfaces when electrostatic interactions dominate. This charge-driven protein orientation hypothesis was verified by our simulations results in this work. It was further confirmed by surface plasmon resonance biosensor and time-of-flight secondary ion mass spectrometry experiments reported elsewhere.     

Interfacial biocatalysis on charged and immobilized substrates: the roles of enzyme and substrate surface charge

An enzyme charge ladder was used to examine the role of electrostatic interactions involved in biocatalysis at the solid-liquid interface. The reactive substrate consisted of an immobilized bovine serum albumin (BSA) multilayer prepared using a layer-by-layer technique. The zeta potential of the BSA substrate and each enzyme variant was measured to determine the absolute charge in solution. Enzyme adsorption and the rate of substrate surface hydrolysis were monitored for the enzyme charge ladder series to provide information regarding the strength of the enzyme-substrate interaction and the rate of interfacial biocatalysis. First, each variant of the charge ladder was examined at pH 8 for various solution ionic strengths. We found that for positively charged variants the adsorption increased with the magnitude of the charge until the surface became saturated. For higher ionic strength solutions, a greater positive enzyme charge was required to induce adsorption. Interestingly, the maximum catalytic rate was not achieved at enzyme saturation but at an invariable intermediate level of adsorption for each ionic strength value. Furthermore, the maximum achievable reaction rate for the charge ladder was larger for higher ionic strength values. We propose that diffusion plays an important role in interfacial biocatalysis, and for strong enzyme-substrate interaction, the rate of diffusion is reduced, leading to a decrease in the overall reaction rate. We investigated the effect of substrate charge by varying the solution pH from 6.1 to 8.7 and by examining multiple ionic strength values for each pH. The same intermediate level of adsorption was found to maximize the overall reaction rate. However, the ionic strength response of the maximum achievable rate was clearly dependent on the pH of the experiment. We propose that this observation is not a direct effect of pH but is caused by the change in substrate surface charge induced by changing the pH. To prove this hypothesis, BSA substrates were chemically modified to reduce the magnitude of the negative charge at pH 8. Chemical modification was accomplished by the amidation of aspartic and glutamic acids to asparagine and glutamine. The ionic strength response of the chemically modified substrate was considerably different than that for the native BSA substrate at an identical pH, consistent with the trend based on substrate surface charge. Consequently, for substrates with a low net surface charge, the maximum achievable catalytic rate of the charge ladder was relatively independent of the solution ionic strength over the range examined; however, at high net substrate surface charge, the maximum rate showed a considerable ionic strength dependence.     

Control of specific attachment of proteins by adsorption of polymer layers

This study concerns the design of protein-resistant polymer adsorbed layers for the control of surface binding of biospecific recognition entities. Polymer surface layers were prepared using the adsorption of poly(allylamine hydrochloride) (PAH), poly(l-lysine) (PL), and branched and linear polyethyleneimine (PEI) and further modified by the covalent attachment of biotin for specific avidin attachment. The adsorption of PAH, PL, and PEI on silicon substrates was studied as a function of pH and ionic strength using ellipsometry. Average dry layer thicknesses of approximately 10, approximately 5, approximately 9, and approximately 3 A (+/-1 A) were obtained when polymer adsorption occurred from solutions at pH 9.5 that contained 0.5 M NaCl for PAH, PL, branched PEI, and linear PEI, respectively. These polymers showed significant differences in their efficiency to suppress nonspecific avidin adsorption. At low ionic strength, avidin adsorption occurred on all polymer-coated surfaces at basic pH values, despite the same positive electrostatic charge for protein globules and the surface. Though the net electrostatic repulsion between avidin molecules and branched PEI was efficiently screened in a protein solution of pH 7 and 0.15 M NaCl, branched-PEI coatings of high molecular weight were unique in their ability to provide avidin-resistant surfaces as a result of steric hindrance from the branched architecture of adsorbed polymer chains. All polymers studied were effective in suppressing avidin adsorption at pH 3 as a result of protonation of the avidin surface functional groups at this pH. Branched-PEI-coated surfaces were also effective for the suppression of smaller positively charged proteins such as lysozyme and ribonuclease A at pH 7 and 0.15 M NaCl. They were also resistant to the adsorption of negatively charged proteins such as BSA and fibrinogen at pH 7 and 0.75 M NaCl. Furthermore, by using PEI-modified protein-repellent surfaces, selective binding of avidin was achieved to surface-bound silver nanoparticles, which should provide a promising application for the label-free detection of biological species using surface-enhanced Raman scattering (SERS).     

Adsorption of monoclonal IgG on polystyrene microspheres

In this paper the adsorption of a monoclonal antibody IgG-1 isotype against HBsAg onto positively and negatively charged polystyrene beads has been studied. To determine the role played by electrostatic forces in the adsorption process different pH values were used. It was confirmed that the affinity of adsorption isotherms depends on the electrostatic interaction between protein and polymer surface. The maximum adsorption amount is located around the i.e.p. of the dissolved protein, and decreases markedly as pH moves away. Thus, the major driving force for adsorption of monoclonal antibodies on polystyrene beads comes from the hydrophobic interaction between the antibody molecules and the adsorbent surface. Desorption of preadsorbed IgG molecules by increasing ionic strength has shown that the positively charged polystyrene is also more hydrophobic in character than the negatively charged one. Finally, electrokinetic experiments have determined that the electric double layer (e.d.l.) of monoclonal antibody changes as the consequence of adsorbing on charged polymer surfaces.     

The role of electrostatic interactions in protease surface diffusion and the consequence for interfacial biocatalysis

This study examines the influence of electrostatic interactions on enzyme surface diffusion and the contribution of diffusion to interfacial biocatalysis. Surface diffusion, adsorption, and reaction were investigated on an immobilized bovine serum albumin (BSA) multilayer substrate over a range of solution ionic strength values. Interfacial charge of the enzyme and substrate surface was maintained by performing the measurements at a fixed pH; therefore, electrostatic interactions were manipulated by changing the ionic strength. The interfacial processes were investigated using a combination of techniques: fluorescence recovery after photobleaching, surface plasmon resonance, and surface plasmon fluorescence spectroscopy. We used an enzyme charge ladder with a net charge ranging from -2 to +4 with respect to the parent to systematically probe the contribution of electrostatics in interfacial enzyme biocatalysis on a charged substrate. The correlation between reaction rate and adsorption was determined for each charge variant within the ladder, each of which displayed a maximum rate at an intermediate surface concentration. Both the maximum reaction rate and adsorption value at which this maximum rate occurs increased in magnitude for the more positive variants. In addition, the specific enzyme activity increased as the level of adsorption decreased, and for the lowest adsorption values, the specific enzyme activity was enhanced compared to the trend at higher surface concentrations. At a fixed level of adsorption, the specific enzyme activity increased with positive enzyme charge; however, this effect offers diminishing returns as the enzyme becomes more highly charged. We examined the effect of electrostatic interactions on surface diffusion. As the binding affinity was reduced by increasing the solution ionic strength, thus weakening electrostatic interaction, the rate of surface diffusion increased considerably. The enhancement in specific activity achieved at the lowest adsorption values is explained by the substantial rise in surface diffusion at high ionic strength due to decreased interactions with the surface. Overall, knowledge of the electrostatic interactions can be used to control surface parameters such as surface concentration and surface diffusion, which intimately correlate with surface biocatalysis. We propose that the maximum reaction rate results from a balance between adsorption and surface diffusion. The above finding suggests enzyme engineering and process design strategies for improving interfacial biocatalysis in industrial, pharmaceutical, and food applications.     

SANS and SAXS analysis of charged nanoparticle adsorption at oil-water interfaces

A systematic study of the adsorption of charged nanoparticles at dispersed oil-in-water emulsion interfaces is presented. The interaction potentials for negatively charged hexadecane droplets with anionic polystyrene latex particles or cationic gold particles are calculated using DLVO theory. Calculations demonstrate that increased ionic strength decreases the decay length of the electrostatic repulsion leading to enhanced particle adsorption. For the case of anionic PS latex particles, the energy barrier for particle adsorption is also reduced when the surface charge is neutralized through changes in pH. Complementary small-angle scattering experiments show that the highest particle adsorption for PS latex occurs at moderate ionic strength and low pH. For cationic gold particles, simple DLVO calculations also explain scattering results showing that the highest particle adsorption occurs at neutral pH due to the electrostatic attraction between oppositely charged surfaces. This work demonstrates that surface charges of particles and oil droplets are critical parameters to consider when engineering particle-stabilized emulsions.     

The Effect of Surface Charge on Adsorption of a Cationic Polyelectrolyte

The regularities of adsorption of a cationic polyelectrolyte, poly(diallyldimethylammonium chloride), on the surface of fused quartz are studied at different values of solution pH by capillary electrokinetics. It is shown that the polyelectrolyte adsorption on a negatively charged surface depends on the value of the surface charge and increases with its growth. At a low charge value (pH 3.8), the polyelectrolyte adsorption increases the quartz surface charge. The driving forces of the adsorption are both electrostatic interaction and forces of nonelectrostatic nature, probably hydrophobic interactions and a change in entropy due to the displacement of counterions from a double layer. The adsorption of poly(diallyldimethylammonium chloride) on quartz from alkaline and neutral solutions is irreversible, which indicates the key role of the electrostatic interaction. At low values of the surface charge, the nonelectrostatic interactions play the main role, thereby resulting in polyelectrolyte desorption.     

The role of variable surface charge and surface complexation in the adsorption of humic acid on magnetite

The ionic strength dependence of humic acid (HA) adsorption on magnetite (Fe₃O₄) was investigated at pH 5, 8 and 9, where variable charged magnetite is positive, neutral and negative, respectively. The adsorption studies revealed that HA has high affinity to magnetite surface especially at lower pH, where interacting partners have opposite charges. However, in spite of electrostatic repulsion at pH 9 notable amounts of humate are adsorbed. Increasing ionic strength enhances HA adsorption at each pH due to charge screening. The dominant interaction is probably a ligand-exchange reaction, nevertheless the Coulombic contribution to the organic matter accumulation on oxide surface is also significant under acidic condition. The results from size exclusion chromatography demonstrate that the smaller size HA fractions enriched with functional groups are adsorbed preferentially on the surface of magnetite at pH 8 in dilute NaCl solution.     

Adsorption of the protein bovine serum albumin in a planar poly(acrylic acid) brush layer as measured by optical reflectometry

The adsorption of bovine serum albumin (BSA) in a planar poly(acrylic acid) (PAA) brush layer has been studied by fixed-angle optical reflectometry. The influence of polymer length, grafting density, and salt concentration is studied as a function of pH. The results are compared with predictions of an analytical polyelectrolyte brush model, which incorporates charge regulation and excluded volume interactions. A maximum in adsorption is found near the point of zero charge (pzc) of the protein. At the maximum, BSA accumulates in a PAA brush to at least 30 vol %. Substantial adsorption continues above the pzc, that is, in the pH range where a net negatively charged protein adsorbs into a negatively charged brush layer, up to a critical pH value. This critical pH value decreases with increasing ionic strength. The adsorbed amount increases strongly with both increasing PAA chain length and increasing grafting density. Experimental data compare well with the analytical model without having to include a nonhomogeneous charge distribution on the protein surface. Instead, charge regulation, which implies that the protein adjusts its charge due to the negative electrostatic potential in the brush, plays an important role in the interpretation of the adsorbed amounts. Together with nonelectrostatic interactions, it explains the significant protein adsorption above the pzc.     

A thermodynamic study of protein-induced lipid lateral phase separation. Effect of lysozyme on mixed lipid vesicles

In this work we study by differential scanning calorimetry (DSC) the lateral phase separation induced by a globular protein (lysozyme) on vesicles built-up by charged (phosphatidic acid) and neutral (phosphatidylcholine) lipids.The adsorption of the positively charged protein onto the negative vesicle surface induces the formation of micro-domains richer in the charged lipid component. This phenomenon is revealed as a splitting of the excess heat capacity peak associated to the melting of the lipid hydrocarbon chains.Also, the peak associated to the protein denaturation is shifted, suggesting the presence of adsorbed proteins onto the vesicle surface. The surface electrostatic potentials, both of proteins and vesicles, have been modulated by pH and ionic strength variations, showing a deep influence of the electric charges in modifying protein adsorption, rate of denaturation (related to unfolding enthalpy variation), and lipid micro-domain formation.Some of the present results have been rationalized on the basis of a theoretical model recently developed by the authors.