阳极电泳和阴极电泳的快速半干新技术

滤纸条半干技术在保证分辨率的前提下, 将凝胶电泳的电泳时间从原来的2～4h缩短到40～50min, 并简化了操作, 节省了实验费用, 不需配制大量的缓冲液. pH4.8较pH8.9阳极电泳提高了酸性蛋白质的电泳分辨率. pH5.5阴极电泳用于分离碱性样品可以得到很好的效果.     

卷柏总蛋白质提取方法及双向电泳条件的建立

目的:建立复苏植物——卷柏总蛋白质的提取方法,以及可以对其蛋白质组进行阵列分离的双向电泳条件。方法:通过多种条件的组合与优化,建立了以物理和化学2种方法充分裂解植物组织细胞,并采用低温操作、加入PVP等去除植物组织中大量的酚类物质,最后离心去除不溶性杂质的卷柏蛋白质组双向电泳(2-DE)样品制备方法。第1向电泳为固相pH梯度等电聚焦,第2向电泳为垂直平板SDS-PAGE。结果:通过对样品制备、蛋白定量、第1向和第2向电泳、染色方法等各实验环节进行控制和优化,凝胶经考马斯亮蓝染色后,可分辨蛋白质斑点数约600个。结论:建立了卷柏总蛋白质的提取方法及蛋白质组双向电泳技术,为后续研究卷柏在干旱胁迫下的差异表达奠定了基础。     

微流控芯片电泳及其法医学应用

在光学性能良好的Brofloat玻璃电泳芯片上,利用自行搭建的共聚焦激光诱导荧光检测系统,通过对芯片管道表面修饰、筛分介质、分离电场强度、进样方式、电泳温度、进样时间等条件的优化,对含15个STR基因座的法医DNA样品进行电泳分离测试实验.通过对芯片电泳条件优化获得了本电泳系统的最佳条件,成功实现8 min内完成DNA样品片段的分离,表明该微流控芯片电泳系统在法医DNA快速分析方面具有良好的应用前景.     

蛋白质检测的方法学研究概述

蛋白质作为生命活动中起重要作用的生物大分子,与一切揭开生命奥秘的重大研究课题都有密切的关系.蛋白质的分离与检测是蛋白质研究的主要技术之一.我们就蛋白质检测的常规方法、电化学检测方法、分子生物学检测方法、电泳法和质谱法等进行了简要综述.     

结肠癌蛋白质双向电泳技术的建立

目的:建立结肠癌13cm和24cm非线性分离系统的2-D图谱,分析比较两者的分辨率.方法:提取结肠癌总蛋白,用pH3-10非线性干胶条对样品进行等电聚焦分离,并分别使用13cm和24cm电泳系统进行双向电泳,考马斯亮蓝G250染色,图像分析,比较对比两组2-D图谱,量化分析两种系统的分辨率差异.结果:在等点电3-10,分子量20-170 kD范围内分别分离得到蛋白质斑点873个(13 cm电泳系统)和1349个(24cm电泳系统).对于24cm电泳系统,1 mg蛋白质上样量的电泳图谱清晰,分辨率较好.结论:成功建立了高分辨率、简便易控的结肠癌蛋白质组双向电泳技术平台.     

蛋白质组研究中分离新技术与新方法

对于蛋白质组的研究离不开分析技术的支撑。由于样品及其基质的复杂性,为了实现蛋白质的高通量、高灵敏度、快速分析鉴定,必须发展与之匹配的新技术与新方法。多维高效液相色谱/毛细管电泳技术,部分弥补了传统2D PAGE的不足,近年来,在蛋白质分离鉴定方面取得了最令人瞩目的成绩。本文分别从多维液相色谱分离技术、多维毛细管电泳蛋白质分离平台、微柱液相-毛细管电泳联用技术、极端pH蛋白质的分离分析和蛋白质的在线富集技术等方面对蛋白质组学研究中在新技术与新方法方面近期取得的成果加以系统阐述。     

连续自由流电泳样机的应用

用连续自由流电泳(continuous flow electrophoresis,CFE)成功地将细胞色素c(Cyt c)和牛血红蛋白(Hb)两种模式蛋白分开,电泳分离后的样品经过紫外分光光度计和聚丙烯酰胺凝胶电泳检测.样品的分离结果与众多性能参数有关,如:分离室间隙,样品流速,缓冲液流量、pH、电导率及分离功耗等.     

大肠癌双向电泳分离条件的建立及其初步分析

采用固相pH梯度等电聚焦为第一向,SDS-PAGE垂直电泳为第二向建立了大肠癌双向电泳分离技术实验条件,对样品的处理、水化、等电聚焦、凝胶平衡等步骤进行了优化,成功地获得了大肠癌分辨率高、重复性好的双向电泳图谱.癌组织样品经3次重复实验共获得蛋白质斑点数1 186±46个,蛋白质斑点位置在IEF方向平均偏差为1.67±0.29 mm,在SDS-PAGE方向为1.41±0.16 mm,蛋白质表达量的相对标准差为6.67 %±2.25 %.经ImageMaster 2D Elite软件初步分析后发现了一些差异表达的蛋白质.     

微芯片——生命科学领域的新工具

微芯片（有人称其为生物芯片biochip)是用硅、玻璃等材料,经光刻、化学合成等技术微加工而成的、大小1 cm²左右的芯片.它可以用来对生物样品进行分离、制备、预浓缩,还可以作为微反应池进行PCR(polymerase chain reaction)、LCR(ligase chain reaction)等反应.最为吸引人的是,芯片上制成多种不同的DNA阵列,即可用于核酸序列的测定及基因突变检测.对微芯片的制作、作用原理、性能及用途等进行了综述.     

蛋白质组分析技术进展

蛋白质组是指某一物种、个体、器官、组织、细胞乃至体液在精确控制其环境条件之下,特定时刻的全部蛋白质表达图谱。继基因组之后,综的研究即将成为分子生物学的研究热点,蛋白质组研究中常用分离分析技术包括样品制备,双向凝胶电泳,毛细管电泳,色谱技术和质谱技术。双向凝胶电泳是在较短时间内分离大量蛋白质组分,提供足够分离空间的比较成熟的方法。各种分离技术的连用和分析过程的自动化将是蛋白质组研究技术的发展方向。     

真空冷冻干燥技术在食用菌加工中的应用研究

简析了食用菌冻干制品产业发展形势,概述了真空冷冻干燥技术原理及产品特点,阐述了应用真空冷冻干燥技术加工食用菌制品所需的主要设备、生产工艺流程及操作要点。探讨真空冷冻干燥过程中,各项工艺参数对食用菌干燥特性的影响。介绍食用菌冷冻干燥技术的优越性、技术壁垒及发展前景,旨在为采用真空冷冻干燥技术对食用菌进行干燥加工提供参考。     

Cell culture processes for monoclonal antibody production

Animal cell culture technology has advanced significantly over the last few decades and is now generally considered a reliable, robust and relatively mature technology. A range of biotherapeutics are currently synthesized using cell culture methods in large scale manufacturing facilities that produce products for both commercial use and clinical studies. The robust implementation of this technology requires optimization of a number of variables, including 1) cell lines capable of synthesizing the required molecules at high productivities that ensure low operating cost; 2) culture media and bioreactor culture conditions that achieve both the requisite productivity and meet product quality specifications; 3) appropriate on-line and off-line sensors capable of providing information that enhances process knowledge; and 4) good understanding of culture performance at different scales to ensure smooth scale-up. Successful implementation also requires appropriate strategies for process development, scale-up and process characterization and validation that enable robust operation that is compliant with current regulations. This review provides an overview of the state-of-the art technology in key aspects of cell culture, e.g., engineering of highly productive cell lines and optimization of cell culture process conditions. We also summarize the current thinking on appropriate process development strategies and process advances that might affect process development.     

Cell culture processes for monoclonal antibody production

Animal cell culture technology has advanced significantly over the last few decades and is now generally considered a reliable, robust and relatively mature technology. A range of biotherapeutics are currently synthesized using cell culture methods in large scale manufacturing facilities that produce products for both commercial use and clinical studies. The robust implementation of this technology requires optimization of a number of variables, including (1) cell lines capable of synthesizing the required molecules at high productivities that ensure low operating cost; (2) culture media and bioreactor culture conditions that achieve both the requisite productivity and meet product quality specifications; (3) appropriate on-line and off-line sensors capable of providing information that enhances process control; and (4) good understanding of culture performance at different scales to ensure smooth scale-up. Successful implementation also requires appropriate strategies for process development, scale-up and process characterization and validation that enable robust operation and ensure compliance with current regulations. This review provides an overview of the state-of-the art technology in key aspects of cell culture, e.g., generation of highly productive cell lines and optimization of cell culture process conditions. We also summarize the current thinking on appropriate process development strategies and process advances that might affect process development.Key words: monoclonal antibody, expression systems, cell line engineering, cell culture process development, optimization scale-up and technology transfer, process advances     

高酸值花椒种籽油的碱炼及工艺

在前人研究的基础上,对高酸值花椒籽油的碱炼除酸实用技术进步进一步的研究。结果表明,传统的常规碱炼技术可以对酸值在２０以下的花椒籽油进行精炼;用采用稀溶液分步间歇式碱炼则可以处理酸值高达６０－７０的花椒籽油。实验室试验表明稀溶液分步间歇式碱炼的工艺流程,可以在低炼耗的水平上得到酸值较理想的花椒籽碱炼油,同时该碱炼方法 显著的降低花椒籽油的色度。     

An overview of continuous protein purification processes

As the sphere of influence of recombinant technology moves away from the laboratory bench, towards product commercialization, development of manufacturing and large scale process technology is becoming a major challenge and determinant for commercial success. The challenge is particularly acute for protein purification process development where protein purification costs tend to dominate overall process economics. The primary objective for process scale purification is to minimize cost for a purified product which meets specifications. Continuous processes may be used to facilitate achievement of the overall objectives. This review critically examines the use of continuous processing for protein purification and recovery operations. The processes have been divided into three general areas: adsorptive and chromatographic, electrophoretic, and extractive. Consideration is given to the operational advantages and limitations of the reviewed processes.     

患者安全事件自动化识别技术的应用

患者安全事件的自动化识别研究作为医疗卫生领域里一项新兴技术,是信息技术革新的产物,拥有光明的发展前景和勃勃的学术生机。介绍了这一技术领域的使用现状和研究进展,并将国内外医疗团队对这一新兴技术项目的研究和使用现状进行了归纳,为医疗机构加强患者安全管理提供参考。     

Key composition optimization of meat processed protein source by vacuum freeze-drying technology

Vacuum freeze-drying technology is a high technology content, a wide range of knowledge of technology in the field of drying technology is involved, it is also a method of the most complex drying equipment, the largest energy consumption, the highest cost of drying method, but due to the particularity of its dry goods: the freeze-drying food has the advantages of complex water performance is good, cooler and luster of freezing and drying food to maintain good products, less nutrient loss, light weight, easy to carry transportation, easy to long-term preservation, and on the quality is far superior to the obvious advantages of other dried food, making it become the forefront of drying technology research and development. The freeze-drying process of Chinese style ham and western Germany fruit tree tenderloin is studied in this paper, their eutectic point, melting point and collapse temperature, freeze-drying curve and its heat and mass transfer characteristics are got, then the precool temperature and the highest limiting temperature of sublimation interface are determined. The effect of system pressure on freeze-dried rate in freeze-drying process is discussed, and the method of regulating pressure circularly is determined.     

Simulation for the recovery of plasmid for a DNA vaccine

The pIDKE2 plasmid is the main component of the CIGB's candidate vaccine against Hepatitis C virus (HVC), which is being used in HCV chronically-infected individuals during clinical trials phase 1 and 2. The designed downstream process of pIDKE2 plasmid produces up to 179 g/year. In order to conduct further improvements, modelling of the downstream process was performed. A methodology based on process analysis tools, such as experimental design and modelling was established to identify factors with the highest influence on production cost and the amount of annual plasmid. Taking into account that the pIDKE2 downstream process designed is in its initial stages of development, CIM technology was evaluated as a new manufacturing process on lab scale. Purity and recovery of CIM technology was better than porous particle matrix, thus SuperPro Designer was used in order to simulate the purification process. Cost efficiency optimization of the pIDKE2 downstream process was done with the simulation model.     

我国医疗技术管理中引入医疗技术评估的路径探讨

2015年以来,在简政放权的大环境下,我国医疗技术管理面临着从重审批的分级分类管理向重监管转型的挑战和机遇,亟需探索建立更加科学合理的医疗技术管理体系。使用医疗技术评估的科学工具和手段是在医疗技术管理领域国际通行的理念和做法。本文梳理了我国医疗技术管理政策沿革,介绍了国际典型代表英国医疗技术管理的过程和方法,并为完善我国医疗技术管理提供了几条政策建议。     

工业生物过程智能控制原理和方法进展

工业生物过程是一个复杂的系统过程,对活体细胞代谢过程的认识是实现高效工业生物制造的基础。文中首先综述了工业发酵过程多尺度优化控制原理和实践,包括多尺度理论与装备、细胞宏观代谢在线检测传感技术以及生理代谢参数相关分析。在此基础上,对工业生物过程智能控制——感知细胞内生理代谢特性新型传感技术、大数据库建立和数据深度计算以及生物过程智能决策进行了综述和展望。