Automated flow-based anion-exchange method for high-throughput isolation and real-time monitoring of RuBisCO in plant extracts

In this work, a miniaturized, completely enclosed multisyringe-flow system is proposed for high-throughput purification of RuBisCO from Triticum aestivum extracts. The automated method capitalizes on the uptake of the target protein at 4 °C onto Q-Sepharose Fast Flow strong anion-exchanger packed in a cylindrical microcolumn (105 × 4 mm) followed by a stepwise ionic-strength gradient elution (0-0.8 mol/L NaCl) to eliminate concomitant extract components and retrieve highly purified RuBisCO. The manifold is furnished downstream with a flow-through diode-array UV/vis spectrophotometer for real-time monitoring of the column effluent at the protein-specific wavelength of 280 nm to detect the elution of RuBisCO. Quantitation of RuBisCO and total soluble proteins in the eluate fractions were undertaken using polyacrylamide gel electrophoresis (PAGE) and the spectrophotometric Bradford assay, respectively. A comprehensive investigation of the effect of distinct concentration gradients on the isolation of RuBisCO and experimental conditions (namely, type of resin, column dimensions and mobile-phase flow rate) upon column capacity and analyte breakthrough was effected. The assembled set-up was aimed to critically ascertain the efficiency of preliminary batchwise pre-treatments of crude plant extracts (viz., polyethylenglycol (PEG) precipitation, ammonium sulphate precipitation and sucrose gradient centrifugation) in terms of RuBisCO purification and absolute recovery prior to automated anion-exchange column separation. Under the optimum physical and chemical conditions, the flow-through column system is able to admit crude plant extracts and gives rise to RuBisCO purification yields better than 75%, which might be increased up to 96 ± 9% with a prior PEG fractionation followed by sucrose gradient step.     

Developing counter current chromatography to meet the needs of pharmaceutical discovery

Experiments have been carried out to evaluate Counter Current Chromatography (CCC) as an alternative purification technique to preparative Reverse Phase High Performance Liquid Chromatography (RP-HPLC) for small molecule pharmaceuticals. The major drawback of CCC is the extensive time required in selecting the solvents to perform the separation. This is equivalent to choosing both the stationary phase and the mobile phase at the same time. In RP-HPLC it is a simple matter of deciding on the gradient, most samples can be purified on a C18 column with a water:acetonitrile gradient. The majority of the initial work was based on a standard test set of commercially available compounds, developed within our group to evaluate the performance of the HPLC apparatus and the column prior to the start of work each day. The work carried out on CCC has shown that the technique offers similar capabilities and can be carried out using similar protocols to RP-HPLC. CCC also has some advantages over RP-HPLC and can be regarded as a valuable addition to the chromatography toolbox.     

宽带泵浦激光的聚焦位置对SRS发生过程的影响

 结合SRS实验,重点研究了聚焦位置对Raman介质中的物理过程、Stokes光的输出特性的影响,比较了SRS阈值和转换效率的实验结果与理论预期。     

离心泵叶轮轴面流道计算新方法

现有的两类一元理论离心泵叶轮水力设计方法存在试凑性工作量大或盖板形状不为简单曲线的缺陷。本文提出了符合一元理论的离心泵叶轮轴面流道过流断面的一种新的分布规律,并给出了按照此分布规律计算叶轮轴面流道过流面积的计算方法和公式。将其与传统的两类设计方法做了比较。     

微流量输液泵

对近几年出现的以声、光、电、磁、热等为基本激发形式的新颖的微流量输液泵的原理、特点作了评述。     

叶顶间隙对离心压气机性能影响的研究

叶顶间隙的研究长期以来一直是叶轮机械研究和设计的热点和难点。本文采用CFD软件研究了进口叶尖相对马赫数分别为0．9、1．15、1．3时,有、无间隙的离心叶轮的气动性能,比较了压比、效率及流量的差别。并分析了用于 100 kW微型燃气轮机的高压比、高转速离心压气机在三种不同尺寸叶顶间隙下的性能,讨论了间隙影响的原因。数值模拟结果表明,叶顶间隙在一定范围内时,间隙主要减弱叶轮的升压能力,基本不影响效率和阻塞流量。     

离心泵叶轮参数对进口回流的影响

本文对不同叶片进口角、叶片数、不同叶顶间隙的开式和闭式离心叶轮共12种叶轮的进口流场和回流发生情况进行了探针的动态测试和可视化观察．弄清了上述叶轮参数对进口回流初始流量和进口流场的影响．     

独立双池受激布里渊散射系统中能量转换效率的研究

利用瞬态包含泵浦抽空的SBS理论模型对独立双池受激布里渊散射系统进行数值模拟，给出了能量转换效率随泵浦功率密度的变化规律。实验验证，理论值与实验结果相吻合。     

单模石英光纤中连续波泵浦SRS谱的演化

利用连续波掺Yb双包层光纤激光器为泵浦源,对单模石英光纤中受激Raman散射谱的形成过程进行了实验研究结果表明,由自发Raman散射向受激Raman散射演化的过程中Stokes谱宽度不断变窄当Stokes波信号功率较强时,在Raman光谱内部会出现能量红移现象,使Stokes光谱峰值相对于泵浦波的频移量从440cm^-1转化到490cm^-1.     

Effecfs of Cross-Correlation Between Real Noise in Transient Process of Single-Mode and Imaginary Parts of Colored Pump Laser

A two-dimensional single-mode laser model with cross-correlation between the real and imaginary parts of the colored quadric pump noise is investigated. A novel laser amplitude Langevin equation is obtained, in which the cross-correlation λp between the real and imaginary parts of the pump noise appears. The mean, variance, and skewness of first-passage-time are calculated. It is shown that the mean, variance, and skewness of first-passage-time are strongly affected by λp.