Effects of urea induced protein conformational changes on ion exchange chromatographic behavior

Urea is widely employed to facilitate protein separations in ion exchange chromatography at various scales. In this work, five model proteins were used to examine the chromatographic effects of protein conformational changes induced by urea in ion exchange chromatography. Linear gradient experiments were carried out at various urea concentrations and the protein secondary and tertiary structures were evaluated by far UV CD and fluorescence measurements, respectively. The results indicated that chromatographic retention times were well correlated with structural changes and that they were more sensitive to tertiary structural change. Steric Mass Action (SMA) isotherm parameters were also examined and the results indicated that urea induced protein conformational changes could affect both the characteristic charge and equilibrium constants in these systems. Dynamic light scattering analysis of changes in protein size due to urea-induced unfolding indicated that the size of the protein was not correlated with SMA parameter changes. These results indicate that while urea-induced structural changes can have a marked effect on protein chromatographic behavior in IEX, this behavior can be quite complicated and protein specific. These differences in protein behavior may provide insight into how these partially unfolded proteins are interacting with the resin material.     

Arsenic speciation by gradient anion exchange narrow bore ion chromatography and high resolution inductively coupled plasma mass spectrometry detection

Based on gradient anion exchange chromatography (AEC), a new strategy in As-speciation was evaluated. A narrow bore chromatographic system with lower flow rates (≤300 μL) well suitable for the low flow requirements of higher efficiency nebulizers was splitless coupled to a high resolution sector field ICP MS. The AEC system takes full advantage of the detector sensitivity allowing more diluted samples (50–100 times) to be injected, delivering substantially less sample matrix to the column and a lower eluent load to the plasma. The unique plasma compatibility of the NH₄NO₃-eluent salt used in this study enabled high linear salt ramps in gradient applications, highly reproducible retention times (±1%) and detection limits in the low ng/L range. The separation conditions were applied on two different polymeric anion-exchangers: a low capacity, weakly hydrophobic material (AS11, Dionex) and a more frequently used higher capacity, higher hydrophobic material (AS7, Dionex). On both columns, As-species (As(III/V), MMA, DMA, AsB) and Cl⁻ were separated in less than nine minutes and co-elution was circumvented by adapting the separation pH to the optimal column selectivity. The key-advantage of the NH₄NO₃-eluent is that it can adopt any separation pH without compromising the eluent strength which is not possible with all other eluents used so far. The influences of chloride and methanol were investigated and found not to affect the chromatographic performance. Column deposits caused strong reversible As(v) adsorption which reduced As(v) to As(III). A corresponding phosphate excess in the injected sample eliminated the adsorption and prevented artefacts in As(v)/As(III) ratios. The method applied to ground water samples provided robust separations and is compatible with any sample preservation procedure.     

Behavior of human serum albumin on strong cation exchange resins: II. Model analysis

Experiments with human serum albumin on a strong cation exchange resin exhibit a peculiar elution pattern: the protein elutes with two peaks in a modifier gradient. This behavior is modeled with a general rate model, where the two elution peaks are modeled with two binding conformations, one of which is at equilibrium conditions, while for the other, the adsorption process is rate limited. Isocratic experiments under nonadsorbing conditions were used to characterize the mass transfer process. The isotherm of both adsorption conformations as well as the kinetic of adsorption and desorption for the second conformation are functions of the modifier concentration. They are evaluated with linear modifier gradient experiments and step experiments with various adsorption times. All experimental features are well reproduced by the proposed modified general rate model.     

ZnCl2改性离子交换树脂的制备及其催化乙醇和乙酸酯化反应性能

 采用离子交换法制备了 ZnCl2 改性的阳离子交换树脂催化剂, 考察了 ZnCl2 浓度、离子交换时间和溶剂等对催化剂催化乙酸和乙醇微波酯化反应性能的影响. 结果表明, 以去离子水为溶剂, 以 0.15%ZnCl2 溶液进行离子交换 30 h, 所得催化剂的性能较好. 采用酸碱滴定法、紫外-可见光谱、傅里叶变换红外光谱、扫描电镜和 N2 吸附脱附等手段对催化剂进行了表征. 结果表明, 改性后的催化剂 H+交换量约为原来的 1.5 倍, 表面酸强度增加; 树脂骨架结构变化不大; 紫外吸收峰发生蓝移, 吸收强度减小; 比表面积略有减小; 催化剂在含水体系中表现出较高的催化活性, 且重复使用性能大大提高.     

高透量 PdAu/陶瓷复合膜的表征与性能评测

 采用化学镀方法在非对称氧化铝管状载体上制备出高性能 PdAu 合金膜. 采用阶段性热处理方法通过 X 射线衍射技术监测 PdAu 膜层合金化进程, 发现在 823 K 下 H2 气氛中厚度为 2 μm, Au 含量为 17.37% 的 PdAu 双金属膜层需要 200 h 达到完全合金化; N2/H2 气氛对 PdAu 合金化速率无显著影响, 但 N2 气氛中合金化后的 PdAu 合金膜表面粒子烧结更为严重. 在 623~823 K 下, 与未进行化学镀 Au 时的纯 Pd 膜相比, PdAu 合金膜的 H2 渗透速率平均增加 1.68 倍, 透氢活化能 (Ea) 降低 38.51%; 当温度低于 573 K 时, PdAu 合金膜的 Ea 由高温时的 8.83 kJ/mol 增加到 15.75 kJ/mol, 此时 PdAu 合金膜的 H2 渗透控速步骤由体相扩散为主的 H2 传质步骤过渡到由表面扩散为主的传质步骤. 通过扫描电镜和能谱联用仪对 PdAu 合金膜的膜厚度和截面组成进行了表征. 关键词     

乙醇水汽重整制氢反应中钯铜合金膜的透氢性能

 采用真空化学镀技术制备了厚度较薄的具有 fcc 结构的 Pd0.9Cu0.1/Al2O3 合金复合膜, 在 250~500 °C, 160~310 kPa 的乙醇水汽重整制氢模拟反应条件下, 考察了不同醇/水比的 H2-Ar-C2H5OH-H2O 混合气体系中该膜的透氢性能. 结果表明, 乙醇和水的存在均相同程度地降低了膜的透氢性能, 但测试条件下 Pd-Cu 合金膜仍保持完整的膜致密性和稳定的相结构. 在 500 °C, 310 kPa, H2-Ar-C2H5OH-H2O (H2:Ar:(C2H5OH+H2O) 体积比为 60:30:10) 混合气中, 膜的最大透氢量为 11.3 m3/(m2?h), H2 回收率大于 62.5%. Pd-Cu 合金膜在乙醇水汽重整制氢反应中具有潜在的应用前景.     

Improving hydrophilicity and protein antifouling of electrospun poly(vinylidenefluoride-hexafluoropropylene) nanofiber membranes

<正>Poly(vinylidenefluoride-hexafluoropropylene)(PVDF-HFP) nanofiber membranes with improved hydrophilicity and protein fouling resistance via surface graft copolymerization of hydrophilic monomers were prepared.The surface modification involves atmospheric pressure glow discharge plasma(APGDP) pretreatment followed by graft copolymerization of poly(ethylene glycol) methyl ether methacrylate(PEGMA).The success of the graft modification with PEGMA on the PVDF-HFP fibrous membrane is ascertained by X-ray photoelectron spectroscopy(XPS) and attenuated total reflectance Fourier transform infrared measurements(ATR-FTIR).The hydrophilic property of the nanofiber membranes is assessed by water contact angle measurements.The results show that the PEGMA grafted PVDF-HFP nanofiber membrane has a water contact angle of 0°compared with the pristine value of 132°.The protein adsorption was effectively reduced after PEGMA grafting on the PVDF-HFP nanofiber membrane surface.The PEGMA polymer grafting density on the PVDF-HFP membrane surface is measured by the gravimetric method,and the filtration performance is characterized by the measurement of water flux.The results indicate that the water flux of the grafted PVDF-HFP fibrous membrane increases significantly with the increase of the PEGMA grafting density.     

Preparation of high density polyethylene/polyethylene-block-poly(ethylene glycol) copolymer blend porous membranes via thermally induced phase separation process and their properties

<正>High density polyethylene(HDPE)/polyethylene-block-poly(ethylene glycol)(PE-b-PEG) blend porous membranes were prepared via thermally induced phase separation(TIPS) process using diphenyl ether(DPE) as diluent.The phase diagrams of HDPE/PE-b-PEG/DPE systems were determined by optical microscopy and differential scanning calorimetry(DSC).By varying the content of PE-b-PEG,the effects of PE-b-PEG copolymer on morphology and crystalline structure of membranes were studied by scanning electron microscopy(SEM) and wide angle X-ray diffraction(WAXD). The chemical compositions of whole membranes and surface layers were characterized by elementary analysis,Fourier transform infrared spectroscopy-attenuated total reflection(FTIR-ATR) and X-ray photoelectron spectroscopy(XPS).Water contact angle,static protein adsorption and water flux experiments were used to evaluate the hydrophilicity,antifouling and water permeation properties of the membranes.It was found that the addition of PE-b-PEG increased the pore size of the obtained blend membranes.In the investigated range of PE-b-PEG content,the PEG blocks could not aggregate into obviously separated domains in membrane matrix.More importantly,PE-b-PEG could not only be retained stably in the membrane matrix during membrane formation,but also enrich at the membrane surface layer.Such stability and surface enrichment of PE-b-PEG endowed the blend membranes with improved hydrophilicity,protein absorption resistance and water permeation properties,which would be substantially beneficial to HDPE membranes for water treatment application.     

Electrospinning of zein/chitosan composite fibrous membranes

<正>Zein/chitosan composite fibrous membranes were fabricated from aqueous ethanol solutions by electrospinning. Poly(vinyl pyrrolidone)(PVP) was introduced to facilitate the electrospinning process of zein/chitosan composites.The asspun zein/chitosan/PVP composite fibrous membranes were characterized by scanning electron microscopy(SEM) and tensile tests.SEM images indicated that increasing zein and PVP concentrations led to an increase in average diameters of the composite fibers.In order to improve stability in wet stage and mechanical properties,the composite fibrous membranes were crosslinked by hexamethylene diisocyanate(HDI).The crosslinked composite fibrous membranes showed slight morphological change after immersion in water for 24 h.Mechanical tests revealed that tensile strength and elongation at break of the composite fibrous membranes were increased after crosslinking,whereas Young's modulus was decreased.     

柚皮苷分子印迹膜的水相制备与识别

以柚皮苷为印迹分子,PEG为致孔剂,在水相中制备了柚皮苷分子印迹壳聚糖膜.分别讨论了交联剂、致孔剂和印迹分子的用量对印迹膜结构和性能的影响.用SEM观察了致孔剂对印迹膜形貌及孔径的影响.紫外吸收光谱分析、柚皮苷在不同体系中的溶解度变化,以及红外光谱分析的结果表明功能聚合物壳聚糖和模板分子柚皮苷间形成了氢键.膜的渗透实验结果表明,在水相中,柚皮苷分子印迹膜能有效地从新橙皮苷和柚皮苷的混合液中分离出印迹分子柚皮苷,选择透过率为11.16%.