Characterization of NSE monoclonal antibodies and establishment of a double-antibody sandwich ELISA assay

通讯作者，E-mail: xtli@ xmu.edu.cn 作者简介: 丁焕弟( 1985 年－ ) ，女，在读硕士，主要从事抗肿瘤单克 隆抗体及诊断试剂盒的研究，E-mail: dinghuandi1125 @163.com。[中文文摘]目的:制备并鉴定NSE(Neuron-specific enolase)单克隆抗体,建立可检测NSE蛋白的双抗夹心ELISA方法。方法:用本实验室已表达纯化的NSE融合蛋白免疫BALB/c小鼠,采用杂交瘤技术制备单克隆抗体。采用WB、IP、IF、IHC等方法对获得的NSE单抗进行鉴定及亚型检测。利用辣根过氧化物酶标记纯化后的NSE单抗,建立一个可检测NSE蛋白的双抗夹心ELISA法。结果:通过分析和鉴定,选定2株可稳定分泌抗NSE抗体的杂交瘤细胞株,效价达4.2×107～6.5×107,亚型为IgG2b。免疫印迹结果显示,该抗体不仅能识别细胞内源NSE蛋白,还能识别分泌到细胞培养上清液中的NSE蛋白,此外还可用于免疫荧光及免疫组化检测。文中所建立的双抗夹心ELISA法,最低检测极限为8.85 ng/ml。结论:成功获得了效价高、灵敏度好及特异性强的NSE单抗,建立了一个双抗体夹心ELISA检测系统,具有良好的临床应用前景。[英文文摘]Objective: Preparation and characterization of monoclonal antibodies against NSE protein，and establishment of a double-antibody sandwich ELISA assay． Methods: BALB/c mice were immunized by using purified recombinant NSE，and monoclonal antibodies were generated by hydridoma technique． These antibodies were characterized with ELISA，Western blot，Immunofluorescent and Immunohistochemical staining． The isotypes of these antibodies were determined with an antibody isotyping kit． With Horseradish Peroxidase labelled NSE monoclonal antibody，we were able to establish a double-antibody sandwich ELISA to detect NSE protein． Results: Two positive hybridoma cell lines were selected for test，the titers of these two monoclonal antibodies could reach 4． 2 × 107 －6. 5 × 107，and their isotypes were IgG2b． Our NSE antibodies could detect not only endogernous NSE protein from cells，but also secreted NSE protein from cells in culture medium by Western blot，in addition，they could be used for immunofluorescent and immunohistochemical staining． The minimum amount of NSE protein could be detected by this double-antibody sandwich ELISA was 8． 85 ng /ml． Conclusion: Our NSE monoclonal antibodies achieved good sensitivity and specificity with high titers，and we established a doubleantibody sandwich ELISA assay which could be used for clinical test in future．福建省科技重点项目(编号项目No.2011Y0050); 厦门市科技计划项目(No.3502Z20123009

本轮粮食价格的大起大落:主要原因及未来走势

本文的主要目标是分析2006～2008年国内外农产品价格的变化和原因以及未来粮食价格走势。研究表明:(1)全球粮食价格在2006～2008年上半年出现大幅上涨,但2008年下半年以来又大幅回落,到2008年12月份国际粮食价格已跌至2007年下半年水平,预测2009年粮食价格还将小幅度下降,但中长期的粮价将现上升趋势;(2)全球能源价格的大幅上涨、生物质液体燃料的扩张、市场投机及一些国家所采取的贸易限制政策等因素是导致过去两年粮食价格大幅上涨的主要原因,而全球金融危机导致的石油价格巨幅下挫和生物质液体燃料产业萎缩是2008年下半年以来全球粮食价格大幅回落的主要原因;(3)我国政府粮价控制政策在稳定国内粮食价格方面发挥了重要作用,但农民没有从过去2年多全球粮价上涨中获得应得的利益,2009年可能还将受粮价下降的冲击

发泡镍对肺冲击伤的防护效果及其机理研究

本研究采用激波管,探讨了发泡镍对肺冲击伤的防护效果和防护机理。结果表明,发泡镍可明显减轻肺冲击伤的程度,降低死亡率。发泡镍的防护作用可能与其降低冲击波超压峰值、延长正压上升时间和缩短正压作用时间有关。结果提示发泡镍是一种较为理想的冲击伤防护材料

Development of serial bio-shock tubes and their application

Objective To design and produce serial shock tubes and further examine their application to experimental studies on blast injury. Methods Bio-medical engineering technique was used for the design and development of the serial shock tubes. One thousand four hundred and fifty nine animals (757 rats, 105 guinea pigs, 335 rabbits, 240 dogs and 22 sheep) were then used to test the wounding effects of the shock tubes. Results Three types of bio-shock tubes, that is, large, medium-and small-scale shock tubes were made in our laboratory. The large-scale shock tube is 39 meters long; the inner diameter of the test section is 1 meter; and the maximum overpressure in the driving section is 10.3 MPa. A negative pressure could be formed by means of the reflected rarefactive wave produced by the end plate. The medium-scale shock tube is 34.5 meters long; the maximum overpressure in the driving section is 22 MPa; the test section is designed to be a knockdown, showing 5 basic types with inner diameter of 77 to 600 millimeters, which could be used for researches on overpressure, explosive decompression, underwater explosion, and so on. The small-scale shock tube is 0.5 meter long with the maximum endured overpressure of 68.6 MPa. Results from animal experiments showed that this set of shock tubes could induce various degrees of systemic or local blast injury in large or small animals. Conclusions This set of bio-shock tubes can approximately simulate typical explosive wave produced by nuclear or charge explosion, and inflict various degrees of blast injury characterized by stability and reproducibility. Therefore, they can meet the needs of blast research on large and small animals

~(27,28)P+~(28)Si总反应截面增强及可能相关的机制

测量了27,28P和相应同中子异位素在28Si靶上的中能反应截面.测得N=12和13同中子异位素的反应截面在Z=15处突然增大.对Z≤14同中子异位素和28P的实验数据结果可以用改进的光学极限近似的Glauber理论很好地描述.28P的反应截面能够用扩大核芯以改进的Glauber理论来解释.但是,用改进光学极限和少体近似的Glauber理论却低估了27P的实验数据.理论分析表明,扩大的核芯加质子晕可能是响应27P+28Si反应截面增强的机制