High resolution protein separations using affinity ultrafiltration with small charged ligands

Although the feasibility of affinity ultrafiltration was demonstrated more than 20 years ago, commercial applications have not developed due to the high cost and practical limitations of the large macroligands needed for highly selective separations. The objective of this study was to examine the use of small charged affinity ligands for protein purification by exploiting electrostatic interactions between the charged complex and an electrically-charged membrane. Experiments were performed using bovine serum albumin and ovalbumin with Cibacron Blue as the affinity ligand. Negatively charged versions of a composite regenerated cellulose membrane were generated by covalent attachment of a sulfonic acid functionality. Binding experiments were used to identify appropriate conditions for protein separations. The selectivity for the separation of BSA and ovalbumin was a function of the solution conditions, Cibacron Blue concentration, and membrane charge, with the addition of Cibacron Blue causing a 30-fold increase in selectivity. A diafiltration process was performed at the optimal conditions, giving a BSA product with a purification factor of more than 90-fold and a yield greater than 90%. These results clearly demonstrate the potential of using a small charged affinity ligand for high resolution protein separations.     

等离子体辅助合成分子筛膜及其催化性能的研究

首次将等离子体技术应用于分子筛膜的制备，研究了以微波等离子体处理基材表面分子筛膜前驱体辅助水热反应合成支撑β型分子筛膜．利用XRD、SEM、XPS、BET、TPD表征了分子筛膜的物相、形貌、孔结构、表面元素组成和表面酸性，并通过甲醇与异丁烯液相反应体系实验考察了分子筛膜的催化性能．结果表明，等离子体处理能有效改善分子筛膜前驱体在基材表面的分散状况，减小了分子筛膜晶体的尺度，使晶体大小均匀，形成的膜致密、牢固．与采用常规方法合成的分子筛膜相比，等离子体辅助合成的分子筛膜对甲醇与异丁烯的反应有更好的催化活件．     

二氧化碳与环氧化物的调节共聚合反应速率

在二氧化碳和环氧化物共聚时加入特定的调节剂，生成有规定的分子量和端基官能度的产物．这种调节聚合的本质是调节剂所含活泼氢与活性中心之间的链转移反应．根据建议的历程，提出了调节共聚速率方程，计算结果与实验一致．     

Characterization of SiO2-P2O5-ZrO2 Sol-Gel/NAFION™ Composite Membranes

Composite membranes were prepared by (a) infiltrating NAFION with SiO₂-P₂O₅-ZrO₂ sol, and (b) recasting a film using NAFION solution containing SiO₂-P₂O₅-ZrO₂ sol. The membranes were characterized by Differential Thermal Analysis and ac-impedance spectroscopy as a function of relative humidity. The influences of the heat treatment (80°C–150°C) and cleaning on the electrical properties were investigated. The incorporation of SiO₂-P₂O₅-ZrO₂ gels into NAFION lead to improvements in its thermal stability and proton conductivity.     

Rate processes on fractals: Theory,simulations, and experiments

Heterogeneous kinetics are shown to differ drastically from homogeneous kinetics. For the elementary reaction A + A products we show that the diffusion-limited reaction rate is proportional tot ^– h[A]² or to [A]^x, whereh=1- d _s/2, X=1+2/d _s =(h-2)(h-1), andd _s is the effective spectral dimension. We note that ford = d _s =1, h =1/2 andX = 3, for percolating clustersd _s = 4/3,h = 1/3 andX = 5/2, while for dust ds <1, 1 >h > 1/2 and >X > 3. Scaling arguments, supercomputer simulations and experiments give a consistent picture. The interplay of energetic and geometric heterogeneity results in fractal-like kinetics and is relevant to excitation fusion experiments in porous membranes, films, and polymeric glasses. However, in isotopic mixed crystals, the geometric fractal nature (percolation clusters) dominates.     

Physico-chemical characterization of nanofiltration membranes.

This study presents a methodology for an in-depth characterization of six representative commercial nanofiltration membranes. Laboratory-made polyethersulfone membranes are included for reference. Besides the physical characterization [molecular weight cut-off (MWCO), surface charge, roughness and hydrophobicity], the membranes are also studied for their chemical composition [attenuated total reflectance Fourier spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS)] and porosity [positron annihilation spectroscopy (PAS)]. The chemical characterization indicates that all membranes are composed of at least two different layers. The presence of an additional third layer is proved and studied for membranes with a polyamide top layer. PAS experiments, in combination with FIB (focused ion beam) images, show that these membranes also have a thinner and a less porous skin layer (upper part of the top layer). In the skin layer, two different pore sizes are observed for all commercial membranes: a pore size of 1.25-1.55 angstroms as well as a pore size of 3.20-3.95 angstroms (both depending on the membrane type). Thus, the pore size distribution in nanofiltration membranes is bimodal, in contrast to the generally accepted log-normal distribution. Although the pore sizes are rather similar for all commercial membranes, their pore volume fraction and hence their porosity differ significantly.     

Sol-gel polyimide-silica composite membrane: gas transport properties

The effect of introduction of silica particles prepared by the sol-gel technique on the gas transport properties of a polyimide film was studied. The sorption and permeation of N₂, O₂, CO₂, H₂ and CH₄ were studied and correlated with morphological changes in the polymer structure. From sorption isotherms, we observed that the composite membrane presents higher solubility coefficients than the polyimide one. The solubility coefficient ratio between the composite and the polyimide is about 1.5–2.0. The isotherms were analyzed in terms of the dual mode sorption. The Henry's coefficient and the Langmuir's affinity and saturation constants were obtained allowing to calculate the Langmuir to Henry concentration ratios as function of the gas pressure. These ratios decrease until zero within a certain pressure range as long as the Langmuir's mode is acting and they are higher for the polyimide membrane when compared with the composite one. This study was completed with calorimetric measurements during the sorption. The gas interaction energy in kJ/mol decreases within the same pressure range as previously described. The measured energies are higher for the polyimide film when compared with the composite one because the polyimide membrane presents a stronger energetic effect caused by a higher Langmuir's contribution. From permeation studies at 3.15⁵ Pa, the composite membrane showed higher permeability coefficients and permselectivities than the polyimide one. All these results were explained, taking into account the difference on the imidization degree of both membranes and the morphological changes which may be induced by the silica nodules in the organic/inorganic interphases.     

Electronic spectral and photophysical properties of some p-phenylenevinylene oligomers in solution and thin films

A comprehensive photophysical and spectroscopic study of a new class of p-phenylenevinylene oligomers (PPV-trimers) possessing different alkyl and alkyloxy sidechain substituents and different end groups (aldehyde, CC, phenylene and anthracene units) was undertaken in solution at room temperature (293 K), low temperature (77 K) and in thin films. The study comprises absorption, emission and triplet–triplet absorption spectra, together with quantitative measurements of quantum yields (fluorescence, intersystem crossing, internal conversion and singlet oxygen formation) and lifetimes. The data allow the determination of rate constants for all decay processes. From these, several conclusions could be drawn. Changing from alkyl to alkyloxy substituents does not change fluorescence and internal conversion yields but decreases the (already small) intersystem crossing yield. The introduction of anthracene at the terminal ends of the PPV-trimers leads to the lowest fluorescence yield reported in this study. Of particular importance is the fact that the fluorescence quantum yields in films are of the same order of magnitude as those in solution, which suggests the potential for use of these oligomers for light-emitting device applications. With one of the alkyloxy derivatives, a more detailed study of the early part of the fluorescence decay was made, and it was found that upon excitation a fast conformational relaxation process of the initially excited oligomer occurs, leading to a more planar conjugation segment.     

Influence of particle diameter distribution on protein recovery in the expanded bed adsorption process

The General Rate model has been developed and solved to describe protein adsorption in an expanded bed. The model takes into account axial and local variation of particle size distribution (PSD), external and intra-particle mass transfer resistances, and dispersion in liquid phase. The influence of PSD on breakthrough profiles has been analysed. The simulation results show that for a significantly high expanded bed the lower part of the breakthrough curve profiles, calculated for local particle size distribution (LPSD) and for axial average particle size distribution (APSD) are very similar. However, the upper part of breakthrough profiles calculated for LPSD approaches inlet concentration much more slowly than those calculated for APSD. The retention times of the lower part of uptake curves calculated with average particle diameter are constantly shorter than those obtained from LPSD. For the calculation of the dynamic capacity (DC), the LPSD can be replaced by APSD for large expanded bed heights. Using breakthrough profiles calculated for average particle size, DC values are constantly underestimated.     

Fine characterization of the ethylene and ethane sorption in poly(amide 12-block-tetramethylenoxide) copolymer/AgBF4 membranes

Ethylene/ethane sorption characteristics were determined for dry Pebax™ (poly(amide 12-block-tetramethylenoxide) copolymer)/AgBF₄ membranes by using an electronic microbalance. The membranes containing 0.7 and 22 wt.% AgBF₄ showed a dual-mode sorption isotherm. The ethane isotherms for all the membranes were of the Henry-type, which is the normal sorption for gases in rubbery polymers. The abnormal presence of Langmuir sorption sites only for ethylene in the rubbery copolymer, never reported sofar, is attributed to the silver-based specific complexation sites. The silver salt which dissolved in limited amounts in the rubbery copolymer had a much smaller Langmuir sorption capacity than the salt that crystallized in the copolymer. The sorption kinetics indicate that the crystallized salt did adsorb slowly ethylene according to a zeroth-order kinetics, but not ethane. The gas uptake kinetics resulting from a step of the pressure surrounding the copolymer exhibited one stage for ethane but two stages for ethylene. For the latter, there was first a fast Fickian sorption stage, then a drift of the zeroth-order sorption of ethylene on salt crystals, which contributes for a large part to the total uptake. The zeroth-order sorption suggests that the sorbed ethylene amount in the second-stage is independent of the crystal-surface coverage. The value of the Fickian diffusion coefficient calculated by fitting the kinetics with a solution of the second Fick’s law was 5 × 10⁻¹² m²/s for both ethylene (the first stage) and ethane, and is typical for small organic compounds in a rubbery material.