毛细管电泳技术在重大疾病特异性蛋白质分析中的应用

人类重大疾病特异性蛋白质的识别、鉴定等是目前生命科学亟待解决的问题。毛细管电泳技术用于蛋白质分析,具有装置简单、分析速度快、试剂和样品消耗少以及有多种分离模式和检测手段等特点。本文综述了毛细管电泳技术在人类重大疾病特异性蛋白质分析中的应用,包括肿瘤疾病、神经退行性疾病和输血性传染病等的特异性蛋白质检测。     

在线HILIC-RP二维液相色谱系统的构建及其评价

自1990年Jorgenson和他的学生Bushey实现了蛋白质的全二维分析以来,二维液相色谱发展迅速,已有多种联用模式出现,如SCX-RP、NP-RP、RP-RP等~([1]).其中,以NP-RP联用模式的正交性最高.由于流动相的兼容性不好,NP-RP二维系统往往采用将第一维溶剂蒸发或第一维采用低流量模式与第二维联用.     

SH3结构域序列集的统计耦联分析

基于蛋白质可折叠性和热稳定性具有长程相关性(即耦联性)的事实,使用改进的统计耦联方法分析了SH3结构域序列集中存在的耦联性信息.结果表明统计耦联方法采用的保守性计能量可以较好地评估SH3结构域序列集的位点保守性,具有高平均耦联能量的位点可以基本上对应一些结构或功能上具有重要意义的位点,对统计耦联数据的一些位点扰动个案分析揭示出SH3结构中包含非紧邻扰动和紧邻扰动模式.统计耦联方法结合聚类重排可以对SH3折叠型序列群体中结构核心与非结构核心的位点进行区分,甚至可以区分其中几个功能相关位点的细节差异.SH3结构域中包含了一系列的扰动模式,不同的扰动模式涉及不同的位点组合;各种扰动模式通过一些共有的扰动位点和扰动响应位点相互影响,并最终决定结构中各位点的耦联响应模式.这些耦联信息对于理解蛋白质的序列与结构和功能的关系,以及设计新的蛋白序列有潜在价值.     

氨基己酸为配基的新型弱阳离子交换/疏水双功能色谱固定相对蛋白质的分离纯化

以硅胶为基质、氨基己酸为配基制备了一种新型弱阳离子交换/疏水（WCX/HIC）双功能混合模式色谱固定相。该固定相配基具有一定的疏水性且含有羧基,在高盐浓度下表现为HIC的性质,可作为HIC固定相使用;在低盐浓度条件下表现为离子交换的性质,可作为WCX固定相使用。分别考察了该介质在WCX和HIC两种模式下对标准蛋白质的分离性能,并与商品柱进行比较。结果表明,所合成的WCX/HIC双功能固定相在WCX和HIC两种模式下对蛋白质均有较高的分离度和选择性,且分离能力与商品柱相当,两种模式下标准蛋白质的质量和活性回收率均大于93%,表明该柱具有“一柱二用”的功能,适于生物大分子的分离纯化。基于此双功能色谱柱构建的在线单柱二维液相色谱（2DLC-1C）可在60 min内实现8种蛋白质的快速分离。在70 min内完成了对蛋清中溶菌酶的二维纯化,纯度可达到98.3%。该技术中一根色谱柱可当作两根色谱柱使用,对蛋白质组学研究和重组蛋白药物的生产具有重要的应用价值。     

分子印迹技术在蛋白质分离分析中的研究进展

蛋白质结构复杂,种类多样,与各种生命活动密切相关。大部分蛋白质在生物体中含量极低,对其分析检测带来极大困难。因此实现复杂生物样品中蛋白质的选择性识别与分离,对实现蛋白质的分离分析意义重大。通过分子印迹技术制备的分子印迹聚合物含有与模板分子大小、形状一致,官能团相互匹配的三维印迹空穴,在蛋白质的选择性识别与分离领域显示出了巨大的发展潜力。但是,由于蛋白质具有尺寸较大、构型易变、结构复杂等特点,分子印迹技术在蛋白质印迹中面临着巨大挑战。该文在介绍几种新型分子印迹技术包括表面印迹、抗原决定基印迹和金属螯合物印迹的基础上,综述了近3年分子印迹技术在蛋白质分离分析方面的应用,并对其发展进行了总结与展望。     

基于水溶性共轭聚合物的蛋白质检测*

近年来,以共轭聚合物作为生物传感元件,在生物大分子（如核酸、蛋白质）特异性识别、检测方面的研究越来越受到人们的关注。共轭聚合物具有强的光捕获能力,具有倍增光学响应性,可用来放大荧光传感信号,大大提高检测的灵敏度,为生物传感器的发展提供了新的传感模式。基于共轭聚合物的新型生物传感器在医疗诊断、环境检测以及国家安全防御等方面具有广泛的应用前景。本文简要介绍了共轭聚合物的荧光信号放大机制以及在蛋白质、酶、抗原－抗体检测方面的应用。最后对共轭聚合物在蛋白质检测方面的未来发展趋势进行了展望。     

蛋白质组学质谱平台肽段可检测性预测研究进展

生物质谱是蛋白质组研究中的核心技术之一,可以实现大规模、高通量的蛋白质定性和定量分析。由于样品和实验过程自身的复杂性,质谱实验的重复性还存在一些问题,肽段鉴定和定量结果有很大的随机性,肽段的质谱检测概率问题在蛋白质组研究中,特别是定量蛋白质组研究中备受关注。本文总结了影响肽段可检测性的重要因素,分析了已经提出的计算预测方法,并对其在实验研究中的应用进行了综述。     

功能化磁性纳米材料在磷酸化肽富集中的应用

蛋白质磷酸化是最重要和最普遍的翻译后修饰之一。基于质谱的技术已成为分析蛋白质磷酸化的重要手段。然而,磷酸化肽固有的低丰度和电离效率以及由非磷酸化肽共存引起的严重抑制使得直接质谱分析仍然是一个挑战。为解决此问题,需在质谱分析前对磷酸化蛋白质进行选择性富集。磁性纳米材料具有良好的磁响应性,可以在外界磁铁的帮助下实现与溶液的迅速分离。功能化磁性纳米材料作为一种新型的分析技术已在蛋白质组学研究中得到广泛的应用。该文就近年来对磁性纳米粒子进行各种功能化修饰以提高其特异性吸附能力的吸附材料在磷酸化肽的富集方面的应用予以综述,并展望了功能化磁性纳米材料在磷酸化肽富集领域的应用前景。     

可视交叉传感阵列对蛋白质及其热变性过程的快速识别

以卟啉及其衍生物和特异性染料为敏感化学元件, 基于交叉响应原理构建了识别蛋白质的可视6×6阵列. 该阵列以颜色差谱图显示其与蛋白质作用呈现的特异性光谱反应, 采用聚类分析、 主成分分析和欧氏距离对图谱进行了分析. 结果表明, 该阵列可以鉴别模式蛋白牛血清白蛋白(BSA)、 牛血红蛋白(BHb)和卵清白蛋白(Ova)及其混合物, 且有望实现定量分析. 此外, 阵列的高敏感性使其不仅能识别天然蛋白质和不同变性程度的蛋白质, 还能对其热变性过程进行可视化实时监控. 该阵列产生的特殊颜色变化与蛋白质的空间构型、 微环境pH值的差异及溶解度有关. 因此, 该方法不仅能实现对蛋白质的快速识别, 为蛋白质热变性机理的研究提供新途径, 而且在临床医学和食品安全等的实时快速检测方面有潜在的应用价值.     

适用于质谱分析的在线固相萃取除盐方法

建立了一个简单、快速、有效的适用于质谱或液相色谱-质谱联用的在线固相萃取(SPE)高通量除盐方法。方法分为单柱和双柱模式,借助于包含双梯度泵(上样泵/分析泵)、自动进样器和配有十通切换阀的柱温箱的高效液相色谱系统,完成样品的自动化在线除盐。单柱模式通过上样泵实现在SPE柱上进样和除盐,被分析物则保留在SPE柱上;除盐完成后,通过阀切换利用分析泵洗脱富集在SPE柱上的被分析物。双柱模式则在单柱模式基础上增加了1根SPE柱,在色谱管理软件控制下2根SPE柱轮流工作,高效率完成样品的在线除盐。该方法在结合质谱分析蛋白质、多肽等领域具有较好的应用前景。     

离子液体选择性萃取生物产品

Imidazolium based room temperature ionic liquids have been used to extract selectively L-tryptophan from fermentation broth. BF4 anion was found to enhance dramatically the partitioning of L-tryptophan into ionic liquid phase from aqueous solutions.     

Speciation and Determination of Trace Amount of Inorganic Arsenic in Water,Environmental and Biological Samples

A new speciation and preconcentration method based on dispersive liquid‐liquid microextraction has been developed for trace amounts of As(III) and As(V) in urine and water samples. At pH 4, As(III) is complexed with ammoniumpyrrolidine dithiocarbamate and extracted into 1‐Hexyl‐3‐methylimidazolium hexafluorophosphate, as an ionic liquid (IL) and As(III) is determined by electrothermal atomic absorption spectrometery (ETAAS). Arsenic(V) in the mixing solution containing As(III) and As(V) was reduced by using KI and ascorbic acid in HCl solution and then the procedure was applied to determination of total arsenic. Arsenic(V) was calculated as the difference between the total arsenic content and As(III) content. The effect of various parameters on the recovery of the arsenic ions has been studied. Under the optimum conditions, the enrichment factor 135 was obtained. The proposed method was successfully applied to the determination of trace amounts of As(III) and As(V) in water and biological samples.     

The Phosporylation of Aniline Derivatives in Biphase Systems

Aniline derivatives were phosphorylated in biphase systems using three methods. A comparative study was performed. The best results were obtained when a solid–liquid system was used. This method is the easiest and lead to higher yields (54–81%).     

液体聚异戊二烯的合成与应用研究进展

固态橡胶具有较高的分子量、强度及伸长率,在工业和日常生活中均得到广泛应用.然而,固态橡胶黏度较高,在应用中往往必须引入添加剂.而液体聚异戊二烯是一种液态橡胶,可以作为固态橡胶的软化剂;得益于其特殊的形态和性能,液体聚异戊二烯可望在多方面得到应用.对液体聚异戊二烯的合成与应用进行了评述.     

液体橡皮泥:属性特征、制备策略及应用探索

液体橡皮泥是指空气环境中被颗粒包裹的以可塑性和复杂形状为特征的液体系统,目前已被成功应用于气体传感、蛋白质分析、光催化等领域,并展现出了很多独特优势。这是一种新兴的软物质体系,与被颗粒包裹的形状为类球形的液体弹珠组成相似,但打破了后者的形状单一性。本文从裸液滴和液体弹珠出发,通过对液体形状和颗粒堵塞问题的分析,梳理了液体橡皮泥技术的建立过程。随后,论述了国内外的研究进展,对不同种类液体橡皮泥的制备、特性及应用进行了概括和分析,重点讨论了单层纳米颗粒结构液体橡皮泥的系列研究。最后,围绕液体橡皮泥的概念内涵、制备方法、特性对比、功能应用等问题进行了总结和探讨,并就未来发展方向和研究思路提出了建议。     

Determination of Alkylphenols in Water by Dispersive Liquid–Liquid Microextraction Based on Solid Formation without a Disperser

Dispersive liquid–liquid microextraction based on solid formation without a disperser combined with high-performance liquid chromatography has been developed for the determination of 4-tert-butylphenol, 4-n-nonylphenol, and 4-tert-octylphenol. This method is rapid, easy, and uses only 10 µL of a low toxicity organic solvent (1-hexadecanethiol) for the extraction solvent and no disperser solvent. The extraction time and centrifugation time require less than 10 min. The linear range was 1–500 ng mL^?1 for 4-tert-butylphenol, 2–1000 ng mL^?1 for 4-tert-octylphenol, and 5–500 ng mL^?1 for 4-n-nonylphenol with r² ≥ 0.9986. The detection limits were between 0.2 and 1.5 ng mL^?1. The recoveries of lake and river water samples were in the range of 79% to 108%, and the relative standard deviations were 5% to 10%.     

Applications of multidirectional reflective light-control films on reflective polymer-dispersed liquid crystal displays for enhancement in image quality at lower viewing angles

In this paper, multidirectional light-control reflective (LCR) films are developed in order to create an active reflective structure that will enhance the image brightness and contrast ratio of reflective dye-doped polymer-dispersed liquid crystal (D-PDLC) displays at lower viewing angles. Advantages of LCR films are that their production is low cost and they require a simple photolithographic fabrication method. The optimum design prism-type light-control reflective film succeeded in minimising the surface scattering effect; thus, the contrast ratio is much enhanced. The symmetric and asymmetric LCR films produced multidirectional scattering that enhances the reflectance at lower viewing angles, which has importance in future display applications. In particular, the prism LCR film has been found to be more influential on the reflectance of D-PDLC films due to multidirectional scattering of light by non-symmetric arrays. The improvement in contrast ratio has been confirmed by the enhancement of optical properties for reflective D-PDLC displays at lower viewing angles below 30°.     

Determination of zearalenone in maize products by vortex‐assisted ionic‐liquid‐based dispersive liquid–liquid microextraction with high‐performance liquid chromatography

A novel method has been developed for the analysis of zearalenone in maize products by vortex‐assisted ionic‐liquid‐based dispersive liquid–liquid microextraction combined with HPLC and fluorescence detection. Maize samples were extracted with methanol/water (80:20, v/v) and the extraction solution was then used as the dispersive solvent in the microextraction procedure. The analyte was rapidly transmitted to a small volume of ionic liquid and was determined by HPLC. Various parameters affecting the recovery of the mycotoxin were investigated, such as the type and volume of the extraction solvent, the type and volume of the dispersive solvent, the pH of the aqueous phase, the salt addition, and the time of vortex and centrifugation. Under the optimal experimental conditions, a good linearity of the analyte was obtained in the range of 1.0–1000.0 μg/L with the correlation coefficient of 0.9998. The limit of detection (S/N = 3) and quantification (S/N = 10) were 0.3 and 1.0 μg/kg, and the mean recoveries ranged from 83.5 to 94.9%, with a relative standard deviation less than 5.0%. The proposed method was demonstrated to be simple, cheap, quick, and highly selective and was successfully applied to the determination of zearalenone in maize products.     

Ionic liquid‐based microwave‐assisted surfactant‐improved dispersive liquid–liquid microextraction and derivatization of aminoglycosides in milk samples

A green and simple method, ionic liquid‐based microwave‐assisted surfactant‐improved dispersive liquid–liquid microextraction and derivatization was developed for the determination of aminoglycosides in milk samples. Nonionic surfactant Triton X‐100 and ionic liquid 1‐hexyl‐3‐methylimidazolium hexafluorophosphate were used as the disperser and extraction solvent, respectively. Extraction, preconcentration, and derivatization of aminoglycosides were carried out in a single step. Several experimental parameters, including type and volume of extraction solvent, type and concentration of surfactant, microwave power and irradiation time, concentration of derivatization reagent, and pH value and volume of buffer were investigated and optimized. Under the optimum experimental conditions, the linearities for determining the analytes were in the range 0.4–10.0 ng/mL for tobramycin, 1.0–25.0 ng/mL for neomycin, and 2.0–50.0 ng/mL for gentamicin, with the correlation coefficients ranging from 0.9991 to 0.9998. The LODs for the analytes were between 0.11 and 0.50 ng/mL. The present method was applied to the analysis of different milk samples, and the recoveries of aminoglycosides obtained were in the range 96.4–105.4% with the RSDs lower than 5.5%. The results showed that the present method was a rapid, convenient, and environmentally friendly method for the determination of aminoglycosides in milk samples.     

有机硅侧链液晶研究进展

综述了国内外有机硅侧链液晶近年来的研究进展，对近晶型、席夫碱型、向列型、胆甾型、偶氮苯型、鱼骨型以及光学非线性型几种主要类型的有机硅侧链液晶进行了详细介绍，并对近年来受到重视的有机硅侧链液晶电流性和离聚物研究进展作了较全面的总结，最后对其应用前景进行了展望。