蛋白质在羟基磷灰石界面吸附的研究

考察了酪蛋白酸钠(sodium caseinate,SC)和乳清分离蛋白(whey protein isolate,WPI)在表面性质不同的3种羟基磷灰石(hydroxyapatite,HA)颗粒上的界面吸附,分析了蛋白质的分子构型和HA颗粒的表面性质等因素对蛋白质在HA界面吸附的影响,重点讨论了SC和WPI肽链上磷酸化丝氨酸基团(phosphorylated serine residues,Ser-P)的数量和分布对吸附差异的影响.通过傅里叶变换红外光谱和表面电位分析发现SC和WPI无法被比表面积较小的HA颗粒有效吸附,但是在有效吸附面积较高的球状纳米HA和棒状微米HA上能够被吸附.Ser-P的存在使得SC在HA界面的吸附量更高、吸附能力更强.Ser-P数量和分布的不同则导致了SC中不同的蛋白组分在HA界面的竞争性吸附:β-酪蛋白在2μmHA界面始终存在优先吸附性;当纳米HA的浓度低于15 mg/mL时,纳米HA界面会优先吸附αs-酪蛋白.     

低温苯甲醇醇解法制备高活性锐钛矿型纳米晶TiO2光催化剂

 以TiCl4 为钛源,采用低温苯甲醇醇解法制备了不同粒径及晶相组成的大表面TiO2纳米晶,利用X射线衍射、热重-差热分析、透射电子显微镜、拉曼光谱、紫外-可见漫反射光谱和N2物理吸附等方法考察了焙烧温度和焙烧时间对其晶相组成、晶粒尺寸、比表面积及孔体积等微结构性质的影响,并以苯酚的光催化降解为模型反应评价了样品的光催化活性. 结果表明,未经任何热处理的TiO2样品即为锐钛矿晶相,控制焙烧温度及焙烧时间可进一步调控样品的粒径、比表面积、晶相结构及表面氧缺位浓度. 经400 ℃焙烧3 h制备的纳米晶TiO2具有最佳的光催化活性,其活性比商用Degussa P-25 TiO2更高.     

氨基双四唑型金属螯合磁性纳米粒子的制备及其对蛋白质的吸附性能

通过4步化学反应对磁性Fe3O4@Si02纳米粒子进行化学修饰,设计和制备了一种N,N’-二(5-四唑亚甲基)胺修饰的金属螯合磁性纳米粒子.用X射线光电子能谱(XPS)、Zeta电位对该新型吸附剂进行了表征.用静态吸附法研究了螯合Cu(Ⅱ)吸附剂对溶菌酶、细胞色素C和α-糜蛋白酶的吸附性能以及溶液pH值、盐浓度、蛋白初始浓度对吸附量的影响.结果表明,吸附剂对蛋白质的吸附主要通过金属配位机理进行,且符合Langmuir吸附模型,对溶菌酶、细胞色素C和α-糜蛋白酶的最大吸附量分别20.0、13.5和17.9 mg/g.此外,将螯合Cu(Ⅱ)吸附剂用于混合蛋白质样品的吸附,发现此吸附剂对混合蛋白质样品中的溶菌酶具有选择性吸附作用,说明此金属螯合吸附剂在蛋白质选择性分离富集中具有一定应用价值.     

基底表面浸润性对磁性载体/蛋白纳米复合物吸附的影响

在玻璃基底上分别生长了疏水的ZnO晶种层和超疏水的ZnO纳米线阵列.在磁场作用下1 min内,磁性载体/牛血清白蛋白纳米复合物在基底表面即可达到吸附饱和值;同时发现,以ZnO纳米线阵列为分离基底时的磁分离效率明显高于玻璃片和ZnO晶种层,且这种差距随着纳米复合物浓度的降低而加大,这与超疏水基底具有较小的固-液接触面积及较大的流体剪切力有关.进一步将目标蛋白拓展至血红蛋白及溶菌酶这2种具有不同性质的典型蛋白,证实了该影响规律具有一定的普适性,超疏水基底在提高蛋白磁分离效率方面比商用玻璃、聚丙烯塑料和疏水基底具有更明显的优势,并对"硬"蛋白具有更佳的磁分离效果.本研究有助于进一步理解界面性质对蛋白吸附行为的影响,并为构建新型高效蛋白分离平台开辟了新思路.     

MoS_2/Na_2Fe_2Ti_6O_(16)的水热制备及吸附性能

采用简单的水热法制备出不同复合比例的MoS_2/Na_2Fe_2Ti_6O_(16)(MoS_2/NFTO)纳米复合材料,并对复合材料的形貌、化学组分、比表面积、孔径分布及Zeta电位进行表征和分析,研究了所得产物对甲基蓝(MB)染料的吸附性能及吸附动力学.结果表明,相对于纯Na_2Fe_2Ti_6O_(16),MoS_2质量分数为70%的MoS_2/NFTO复合材料的单位比表面积的吸附容量提高了58倍,对MB的吸附率提高了4.9倍.研究还发现,吸附剂对MB的吸附动力学符合准二级动力学模型.     

含水核载牛血清白蛋白聚氰基丙烯酸丁酯微囊的制备及体外释放

利用界面乳液聚合方法制备了新型含水核载牛血清白蛋白 (BSA)的聚氰基丙烯酸丁酯 (PBCA)纳米微囊 .分别研究了纳米微囊的粒径及其分布 ,表面Zeta电势的变化 .并以牛血清白蛋白为模型药物考察了药物包裹率和载药量的变化以及载药纳米微囊在磷酸缓冲溶液中的体外释放行为 .结果表明 ,所制备的纳米微囊平均粒径为 2 0 0nm ,多分散度为 0 2 2 6;表面Zeta电势的变化证明了BSA是包裹于纳米微囊的内部而不是吸附在其表面 ;包裹率和载药量取决于水相中BSA的初始浓度 ,当BSA的浓度为 0 8mg mL时 ,包裹率和载药量分别为 3 5 %和 0 485× 1 0 - 9mol mg;药物的释放速率取决于纳米微囊的壁厚 ,通过调节壁厚可以达到控释的目的     

应用纳米球刻蚀法在自组装膜修饰的硅表面生成中尺度的网状蛋白层（英文）

在自组装膜修饰的硅表面制备有序的蛋白阵列是研发生物传感器的先决条件之一,因此如何产生有序的表面蛋白阵列一直是生物医药研究方向的前沿。本研究通过应用纳米球刻蚀法在氧化的10-烯基十一烷基三氯硅烷自组装膜修饰的硅表面生成了网状结构溶菌酶蛋白层。网孔的大小(从纳米到微米级别)由表面沉积的纳米球的尺寸来调控。我们利用原子力显微镜和荧光显微镜对样品表面进行了详细表征。结果表明:这种新方法比传统的通过扫描探针在固体表面修饰而聚集溶菌酶蛋白的方法更快捷简便,而且它能够在相对大的硅表面形成网状蛋白层。此外,网孔表面附着具有强吸附活性的羧酸基团层,它可以通过静电吸引或者共价结合来吸附液相中的第二种蛋白分子。     

CeO2-TiO2催化剂的表面结构及其湿式氧化活性

 采用溶胶-凝胶法和共沉淀法制备了CeO2-TiO2催化剂,利用N2吸附、X射线衍射、透射电镜、X射线光电子能谱和Zeta电位分析等手段表征了催化剂的表面结构性质. 以乙酸和苯酚为目标物,进行了湿式氧化高浓度乙酸和苯酚的活性测试,研究了CeO2-TiO2催化剂表面结构与活性之间的关系. 结果表明, Ce和Ti之间的相互作用使复合氧化物CeO2-TiO2催化剂具有小的晶粒尺寸和高的比表面积,催化剂表面化学吸附氧含量增加,且不同制备方法得到的CeO2-TiO2催化剂表面电位点不同,共沉淀法制备的催化剂更有利于乙酸和苯酚的吸附. 在湿式氧化反应中, CeO2-TiO2催化剂有较高的活性,其中共沉淀法制备的催化剂活性最高; 在230 ℃, 5 MPa条件下反应120 min后,乙酸COD去除率为79%; 在150 ℃, 4 MPa下反应120 min后,苯酚COD去除率为96%.     

具特定晶面且大比表面积贵金属纳米晶催化基元的构筑

贵金属纳米晶在电催化等领域具有广泛应用. 其催化活性往往与纳米晶体的表面结构直接相关,而催化剂的贵金属原子利用率与比表面积密切相关. 因小尺寸纳米晶难以保留特定的晶面,而具有特定表面的纳米晶通常结晶成尺寸较大、比表面积比较小的晶体,调控纳米晶的尺寸和表面结构两种策略似乎相互矛盾. 如何可控合成同时具有特定表面结构和大比表面积的贵金属纳米晶具有重要的意义. 本综述从形貌调控角度详细介绍提高贵金属纳米晶原子利用率的方法策略;总结调控单贵金属及其合金同时具有特定晶面和大比表面积的研究现状;最后,对纳米晶的形貌调控领域未来的发展趋势提出展望.     

温敏性丙烯腈共聚物纳米纤维膜的制备与性能

采用可逆加成断裂链转移可控/活性聚合方法合成了丙烯腈与N-异丙基丙烯酰胺(NIPAM)的嵌段共聚物,通过调控嵌段聚合反应时间可以获得一系列不同嵌段链长的共聚物,分子量分布在1.3左右.运用静电纺丝技术制备了所合成嵌段共聚物的纳米纤维膜,扫描电镜照片表明纳米纤维膜较为均匀且直径可调.研究了纳米纤维膜表面水接触角与荧光标记牛血清清蛋白的吸附现象,接触角结果证实共聚物纳米纤维膜具有一定的温度响应性,且疏水性嵌段的引入导致响应温度较PNIPAM有所降低;蛋白质吸附结果则表明温度较低时纳米纤维膜表面更亲水,蛋白质吸附较少.所制备的温敏性纳米纤维膜可望用作智能分离与吸附材料.     

Protein adsorption behaviors onto photocatalytic Ti(IV)-doped calcium hydroxyapatite particles

The fundamental experiments on the adsorption behaviors of proteins onto photocatalytic Ti(4+)-doped calcium hydroxyapatite (TiHap) particles were examined comparing to those onto the calcium hydroxyapatite (CaHap) and commercially available typical titanium oxide (TiO(2)) photocatalyst (TKP-101). The heat treated TiHap and CaHap particles were also used after treated these particles at 650°C for 1h (abbreviated as TiHap650 and CaHap650, respectively). All the adsorption isotherms of bovine serum albumin (BSA), myoglobin (MGB) and lysozyme (LSZ) from 1×10(-4)mol/dm(3) KCl solution were the Langmuirian type. The saturated amounts of adsorbed BSA (n(s)(BSA)) for the CaHap650 particles was higher than that for CaHap. Similar results were observed for TiHap and TiHap650. The adsorption of LSZ exhibited the same result of BSA, while the saturated amounts of adsorbed LSZ (n(s)(LSZ)) value on the TiHap were much higher than CaHap. However, the saturated amounts of adsorbed MGB (n(s)(MGB)) are almost equal to those for the CaHap and TiHap nevertheless whether these particles were heat treated at 650°C or not. The TKP-101 exhibited extremely small adsorption capacity of all proteins due to its small particle size of ca. 4nm in diameter. The independence of the n(s)(MGB) value on the zeta potential (zp) of the particles was explained by the electrostatical neutrality of MGB molecules. On the other hand, the n(s)(LSZ) values were increased with increase in the negative zp of the particles. This fact was explained by increasing the electrostatic attractive forces between negatively charged particles and positively charged LSZ. However, the n(s)(BSA) values exhibit maxima for the heat treated TiHap650 and CaHap650 particles. This result was interpreted to the formation of β-TCP crystal phase by the heat treatment. The produced Ca(2+) ions by dissolution from β-TCP phase may exert as binders between BSA and surfaces of the heat treated particles.     

Adsorption of immunogamma globulin onto various synthetic calcium hydroxyapatite particles

This paper presents data on adsorption of immunogamma globulin (IgG) onto synthetic rodlike calcium hydroxyapatite particles (CaHaps) with various particle lengths and calcium/phosphate (Ca/P) atomic ratios ranging from 1.54 to 1.65 and compares the obtained results to those of acidic (bovine serum albumin, BSA), neutral (myoglobin, MGB), and basic (lysozyme, LSZ) proteins reported before. The effect of electrolyte concentration on IgG adsorption was also examined. The initial rate of IgG adsorption was similar to that of BSA and was slower than that of MGB and LSZ. This fact was interpreted by the difference in the structural stability and molecular weight of these proteins. The isotherms of IgG adsorption onto the CaHap particles were of pseudo-Langmuir type. The saturated amount of adsorbed IgG values (nsIgG) for the particles with mean particle length less than 70 nm decreased with increasing Ca/P ratio. The adsorption behavior of IgG molecules was very similar to that of basic LSZ, though IgG has zero net charge. The nsIgG value was increased with increased mean particle length of CaHaps; the relationship was less significant than that for BSA but similar to those for MGB and LSZ. The similar adsorption behavior of IgG and LSZ suggested that the Fab parts of IgG molecules preferentially adsorb onto CaHap to provide the reversed Y-shaped conformation of IgG. The change of the adsorption mode of IgG molecules from the reversed Y-shaped conformation to side-on by "spreading" the Fc part of IgG molecules onto the particle surface over a longer adsorption time was suggested. The nsIgG value was increased with increasing electrolyte concentration by screening the intra- and intermolecular electrostatic interactions of proteins.     

Protein adsorption characteristics of calcium hydroxyapatites modified with pyrophosphoric acids

Protein adsorption characteristics of calcium hydroxyapatite (Hap) modified with pyrophosphoric acids (PP(a)) were examined. The PP(a) modified Hap particles (abbreviated as PP-Hap) possessed anchored polyphosphate (PP: P-{O-PO(OH)}(n)-OH) branches on their surfaces. The proteins of bovine serum albumin (BSA: isoelectric point (iep)=4.7, molecular mass (M(s))=67,200 Da, acidic protein), myoglobin (MGB: iep=7.0, M(s)=17,800 Da, neutral protein), and lysozyme (LSZ: iep=11.1, M(s)=14,600 Da, basic protein) were examined. The zeta potential (zp) of PP-Hap particles as a function of pH overlapped; zp-pH curves were independent of the concentration of pyrophosphoric acids (abbreviated as [PP(a)]) used for modifying Hap surface. The saturated amounts of adsorbed BSA (Delta n(ads)(BSA)) were increased three-fold by the surface modification with PP(a) though they were independent of the [PP(a)]. Furthermore, the fraction of BSA desorption was independent of the [PP(a)]. This enhancement of BSA adsorption onto the PP-Hap is due to the hydrogen bonding between oxygen and OH groups of the PP-branches and functional groups of BSA molecules. In the case of LSZ, a more higher adsorption enhancement was observed; the saturated amount of adsorbed LSZ (Delta n(ads)(LSZ)) for Hap modified at [PP(a)]=6 mmol/dm(3) was nine-fold than that for Hap unmodified. This remarkable adsorption enhancement was explained by a three-dimensional binding mechanism; LSZ molecules were trapped inside of the PP-branches. Hence, a fraction of LSZ desorption was decreased with an increase in the [PP(a)]; as more PP-branches are presented on the surface the higher retardation of LSZ desorption was induced. It was expected from their small size that MGB adsorb between the PP-branches as well as LSZ. However, the amounts of adsorbed MGB (Delta n(ads)(MGB)) did not vary and were independent of the [PP(a)] due to the small numbers of functional groups of MGB. In addition, no dependence of the fraction of MGB desorption on the [PP(a)] was observed. The results of zp for all the protein systems supported the mode of protein adsorption discussed. The anchored structure of the PP-branches developed on the Hap surface to provide three-dimensional protein adsorption spaces was proved by a comparative experiment that was elucidating the effect of pyrophosphate ions for BSA adsorption onto Hap.     

Adsorption of Bovine Serum Albumin and Lysozyme on Hydrophobic Calcium Hydroxyapatites

The adsorption of bovine serum albumin (BSA) and lysozyme (LSZ) to oleyl phosphate(OP)-grafted calcium hydroxyapatite (OP-CaHAP) with different degrees of hydrophobicity, ranging the number of surface oleyl group per unit nm2 (nO) from 0 to 2.60, was investigated. The pronounced effects of the hydrophobic moiety of adsorbent on protein adsorption were observed. The saturated amount of adsorbed BSA (ns) was increased up to nO = 0.6 by an enlargement of hydrophobic interaction between hydrophobic CaHAP particle and proteins. However, ns decreased at nO >/= 1.3 by increasing the electrostatic repulsive force between negatively charged BSA and OP-CaHAP particles. On the other hand, the ns value of LSZ was continuously increased up to nO = 2.0 and saturated by increasing either the hydrophobic interaction or the electrostatic attraction of positively charged LSZ and negatively charged OP-grafted CaHAPs. The BSA adsorption experiment revealed that the effect of positively charged adsorption sites on the exposed ac or bc crystal faces (C-sites) of the CaHAPs is screened by the OP-groups grafted on their particle surfaces. Copyright 1999 Academic Press.     

Preparation and characterization of calcium phosphate–albumin colloidal particles by high ultrasonic irradiation

Using high intensity ultrasonic irradiation, we prepared calcium phosphate–albumin colloidal particles from aqueous solutions of Ca(H₂PO₄)₂ and Ca(OH)₂ in the presence of bovine serum albumin (BSA). The effect of concentration of BSA (2–5 g/L) properties of the colloidal particles was studied at constant temperature. The effect of a resting period on the size distribution of the colloidal particles was also investigated. Morphology, phase composition, average diameter, size distribution and zeta potential were obtained by transmission electron microscopy, X-ray diffraction, particle size determination by PCS and electrokinetic measurements.     

Evaluation of the adsorption affinity of proteins to calcium hydroxyapatites by desorption and pre-adsorption methods

The adsorption affinity of bovine serum albumin (BSA) and lysozyme (LSZ) to calcium hydroxyapatite (CaHAP) was evaluated by desorption and two step adsorption methods. These experiments were carried out at 15°C in a 1×10⁻⁴ mol dm⁻³ KCl solution of pH 6.0. BSA molecules were scarcely desorbed, exhibiting an irreversible adsorption of BSA, though LSZ slightly desorbed. This result supports our previous findings that LSZ adsorbs weakly onto phosphate ions exposed on ac or bc faces of CaHAP while BSA adsorbs strongly onto positively charged sites on ac or bc faces of CaHAP. The amount of adsorbed LSZ was markedly increased by the pre-adsorption of BSA, where LSZ was adsorbed onto BSA-covered CaHAP. On the other hand, the amount of adsorbed BSA was not changed by the pre-adsorption of LSZ. In both pre-adsorption systems it was confirmed by an HPLC method that no protein molecule pre-adsorbed was desorbed after the post-adsorption procedure. Therefore, it was interpreted that the enhancement of adsorption of positively charged LSZ is induced by an electrostatic attractive force through pre-adsorption of negatively charged BSA molecules with a high coverage. However, since the coverage of LSZ onto CaHAP is considerably low, no stimulation of BSA adsorption occurred on the LSZ-covered surface. The formation of double protein adsorbed layers consisting of pre- and post-adsorbed proteins was proposed.     

Effects of pyrophosphate ions on protein adsorption onto calcium hydroxyapatite

The effects of pyrophosphate ions (PP: P2O7(4-)) on the adsorption of proteins onto calcium hydroxyapatite (Hap) were examined using typical proteins of bovine serum albumin (BSA: isoelectric point (iep) = 4.7, molecular mass (M(s)) = 67 200 Da, acidic protein), myoglobin (MGB: iep = 7.0, M(s) = 17 800 Da, neutral protein), and lysozyme (LSZ: iep = 11.1, M(s) = 14,600 Da, basic protein). The UV and CD measurements determined that both the secondary and the tertiary structures of protein molecules do not vary in the presence of PP. The adsorption of BSA was strongly depressed by the addition of PP in all the methods with changing the order of PP addition. Even if BSA was pre-adsorbed on the Hap surface, PP replaced BSA molecules by strong preferential adsorption onto Hap to reduce the amounts of adsorbed BSA. A similar effect was observed with the adsorption of MGB. On the other hand, the amount of adsorbed LSZ (n(LSZ)) was increased with an increase in the concentration of PP, and the n(LSZ) value showed a maximum point in each adsorption isotherm. This fact was explained by a compression of the electric double layer (EDL) around each LSZ molecule by PP. This compression of the EDL induced the reduction of lateral electrostatic repulsions between charged LSZ molecules on the Hap surface and enhanced the formation of closed-packed monolayers to raise the n(LSZ) value. However, since the number of PPs around a LSZ molecule is decreased by an increase in the LSZ concentration in each system, the thickness of the EDL may be increased. Hence, n(LSZ) was reduced again after the maximum point in each system. Tripolyphosphate (TPP: P3O10(5-)) ions exhibited similar effects on the adsorption behaviors of all proteins, but a much more pronounced effect was observed on the LSZ system. TPP with a higher eletronegativity shielded the EDL more highly than PP to increase the n(LSZ) value. The results of the zeta potential for all the protein systems supported the modes of protein adsorption discussed.     

Selective decomposition of proteins by photocatalytic Ti(IV)-doped calcium hydroxyapatite particles from mixed-protein systems

The decomposition of protein molecules from a mixed-protein solution on the surface of calcium hydroxyapatite (CaHap) and Ti(IV)-doped CaHap (TiHap) particles with a Ti/(Ca + Ti) atomic ratio (X _Ti) of 0.10 and 0.20 under UV irradiation of 365 nm in wavelength was investigated. Acidic bovine serum albumin (BSA) and basic lysozyme (LSZ) were employed as a model of pathogenic proteins. The photocatalytic activities of TiHap particles were estimated from the decomposition of BSA and LSZ from the BSA (2.5 mg/cm³)–LSZ(1.0 mg/cm³) mixture under 1 mW/cm² UV irradiation dispersed in a 10-mL quartz tube. No change in BSA concentration by UV irradiation was observed for all the unheated original CaHap and TiHap particles without and with low photocatalytic activities, respectively. Similar results were observed for the systems that employed heat-treated particles endowed a high photocatalytic activity by heat treatment at 650 °C for 1 h. On the other hand, a selective photocatalytic decomposition was observed for the LSZ, i.e., only LSZ molecules were decomposed completely from the BSA (2.5 mg/cm³)–LSZ(1.0 mg/cm³) mixture by using heat-treated TiHap particles with X _Ti?=?0.10 and 0.20. This selective decomposition by TiHap particles was interpreted by higher adsorption affinity of positively charged LSZ to highly negatively charged TiHap together with low molecular weight and rigid structure of LSZ molecules.     

BSA adsorption on differently charged polystyrene nanoparticles using isothermal titration calorimetry and the influence on cellular uptake

BSA adsorption onto negatively and positively charged polystyrene nanoparticles was investigated. The nanoparticles were characterized in terms of particle size, zeta potential, surface group density, and morphology. The adsorption behavior of BSA on the particle surface, as a function of pH and overall charge of the particle, was studied using ITC. Different thermodynamic data such as enthalpy changes upon binding and stoichiometry of the systems were determined and discussed. The degree of surface coverage with BSA was calculated using the thermodynamic data. The cellular uptake of particles before and after BSA adsorption was studied using HeLa cells in the presence and absence of supplemented FCS in the cell culture medium.     

Selective and sequential adsorption of bovine serum albumin and lysozyme from a binary mixture on nanosized magnetic particles

Magnetic particles about 10 nm in size were prepared by chemical precipitation under nitrogen and used for the selective and sequential adsorption of bovine serum albumin (BSA) (pI = 4.7) and lysozyme (LSZ) (pI = 1.1) under different conditions, such as pH and initial protein concentration. The separation ratio of BSA over LSZ at pH 4.6 is about 5, which is about 1.5 times the separation ratio of LSZ over BSA at pH 11.0. Only 10% of the preadsorbed BSA could be displaced by the sequential adsorption of LSZ at pH 11.0. On the other hand, 60% of the preadsorbed LSZ was desorbed due to the sequential adsorption of BSA at pH 4.6. Over 50% desorption of BSA or LSZ could be achieved either by 0.5 M Na(2)HPO(4) or 0.5 M NaH(2)PO(4) after 2 h. Over 80% of the enzymatic activity of LSZ was preserved when it was desorbed from magnetic particles.