胆固醇配基色谱在蛋白质复性中的应用

以三聚氯氰为连接臂,采用两步法将疏水配基胆固醇修饰到交联琼脂糖（Sepharose）介质上,构建了胆固醇配基色谱。以溶菌酶（Lys）为模型蛋白,考察了胆固醇配基色谱的复性效果,并研究了流速、上样质量浓度和上样量对色谱复性效果的影响,确定了溶菌酶复性的最佳方法。研究结果表明,当溶菌酶上样量为1.00 mL,介质含0.25 mg溶菌酶时,在0.02 mL/min操作流速下进行吸附-洗脱复性,其最终蛋白收率和活性收率分别为96.3%和91.4%,有效促进了溶菌酶的复性。     

干柱层析法分离中药藤黄中的新藤黄酸

目的：分离中药藤黄中的新藤黄酸。方法：采用薄层硅胶G为固定相的干层析柱,以三氯甲烷-甲醇-二乙胺（10：1：1）为流动相进行洗脱,得到一黄色结晶。结果：采用UV,IR,MS,1HNMR和13CNMR方法获得该化合物波谱数据,经与文献报道数据对比,确定所分离得到的化合物为新藤黄酸。结论：干柱层析法可以快速有效地分离中药藤黄中的新藤黄酸。     

大口径毛细管气相色谱法测定萘乙酸

采用大口径毛细管色谱柱,直接进样,建立了一种萘乙酸的气相色谱定量分析新方法,方法的相对标准偏差为０．４１％,回收率在９８．９％～１０１．２％之间。该法操作简便、快速、准确,重现性好。     

固定化金属离子亲和层析研究进展

固定化金属离子亲和层析(IMAC)作为一种新的亲和层析技术在药物,尤其是生物药物制备上已得到越来越广泛的应用。文章综述了固定化金属离子亲和层析的基本原理、构成及应用现状,对其应用的前景进行了展望。     

纸色谱和柱色谱实验方法的改良

对纸色谱和柱色谱实验的方法进行了改良,改良后的实验具有操作方便、现象明显、实验时间短、经济、环保的优点。     

Purification of Stearidonic Acid from Modified Soybean Oil by Argentation Silica Gel Column Chromatography

The objective of this study was to purify stearidonic acid (SDA, 18:4 ω-3) from modified soybean oil containing a mixture of over 20 fatty acids (23% SDA). Interest in obtaining purified fractions of SDA arises from reported health benefits associated with polyunsaturated fatty acids (PUFA), such as cardiovascular disease prevention. In addition, SDA may also provide improved stability characteristics since its unsaturation index is less than longer PUFA, such as eicosapentaenoic acid (EPA, 20:5 ω-3). First, a chemical ethanolysis of modified soybean oil was performed to transform the triacylglycerols into fatty acid ethyl esters (FAEE). Then, the FAEE were fractionated and SDA-EE was purified by argentation silica gel (10% AgNO₃) open column chromatography, which allows selectivity based on degree of unsaturation. Different FAEE sample loads and mobile phases were explored until the best purification of SDA-EE was achieved. The solvents used were hexane and hexane:acetone mixtures (99 and 95%). Under the optimal conditions, a fraction with high SDA-EE purity (96%) was obtained with 77% yield. Besides, it was possible to obtain another fraction enriched in α-linolenic acid-EE (37% purity and 68% yield) and γ-linolenic acid-EE (22% purity and 61% yield). A scaled-up process resulted in 840 mg of final product composed of 97% SDA-EE with 71% yield.     

Gamma-Aminobutyric Acid Production Using Immobilized Glutamate Decarboxylase Followed by Downstream Processing with Cation Exchange Chromatography

We have developed a gamma-aminobutyric acid (GABA) production technique using his-tag mediated immobilization of Escherichia coli-derived glutamate decarboxylase (GAD), an enzyme that catalyzes the conversion of glutamate to GABA. The GAD was obtained at 1.43 g/L from GAD-overexpressed E. coli fermentation and consisted of 59.7% monomer, 29.2% dimer and 2.3% tetramer with a 97.6% soluble form of the total GAD. The harvested GAD was immobilized to metal affinity gel with an immobilization yield of 92%. Based on an investigation of specific enzyme activity and reaction characteristics, glutamic acid (GA) was chosen over monosodium glutamate (MSG) as a substrate for immobilized GAD, resulting in conversion of 2.17 M GABA in a 1 L reactor within 100 min. The immobilized enzymes retained 58.1% of their initial activities after ten consecutive uses. By using cation exchange chromatography followed by enzymatic conversion, GABA was separated from the residual substrate and leached GAD. As a consequence, the glutamic acid was mostly removed with no detectable GAD, while 91.2% of GABA was yielded in the purification step.     

核苷酸的电解致酸脱保护研究

对电致酸脱保护寡核苷酸原位合成体系的溶液性质进行了研究．确定了电解致酸脱保护体系的优化溶液组成,发现选择弱酸HW、电解电位2V和0．1moL／L高氯酸四乙基胺可获得理想的实验结果。     

Estimation of Docosahexaenoic Acid Glycerides in Schizochytrium sp. Oil by Open Column Chromatography and Reversed-phase HPLC

Column chromatography and reversed-phase HPLC using a Sepax BR-C18 column were applied for the analysis of glycerides in the docosahexaenoic acid (DHA) oil by Schizochytrium sp. The oil was detected under UV light after isocratic elution with 20 % 2-propanol/hexane (5:4, v/v) + 80 % acetonitrile. The correlation coefficients for the calibration of HPLC for DHA glycerides were in the range of 0.9958–0.9998. The accuracy was confirmed with recoveries of 93.47–102.25 %. DHA oil samples under different aeration rates were successfully analyzed and the content of the binding of DHA on the acylglycerol backbone was very low (1.81–4.21 %). However, the content of glycerides (tri-, di-, mono-) separated by column chromatography was more than 87.22 %. The glycerides were largely composed of triacylglycerides (TAG) (82.29–89.54 %), where higher content of TAG correlated to higher contents of DHA and TAG of DHA (TriDHA). In addition, the optimal aeration rate of 250 m³/h was obtained for TAG, DHA, and TriDHA production by Schizochytrium sp. with the temperature 30 ± 0.5 °C, impeller speed 85 ± 5 rpm and pressure 0.04 Mpa.     

对脱氢氰酸塔聚合问题的分析

本文对抚顺引进的5万吨/年丙烯腈装置中的脱氢氰酸塔在试生产阶段发生的聚合问题予以介绍,并对聚合产生的原因及延续方法等问题做了分析。     

姜黄色素的简易提取与柱色谱分离研究

利用丙酮-甲苯混合溶剂从姜黄中提取姜黄色素及姜黄油树脂.以体积比为38:1:1的氯仿-乙酸乙酯-甲醇混合溶剂作为洗脱剂,硅胶柱色谱分离10 g姜黄色素,纯物质的收率为:姜黄素45 %,脱甲氧基姜黄素为13 %,二脱甲氧基姜黄素为24 %,纯色素的总收率为82 %.     

反义核酸药物与抗肿瘤研究

现在各国正在利用反义技术发展一类由核酸组成的新型药物,即反义核酸药物。我们合成了5种血管内皮生长因子(VEGF)的反义寡聚脱氧核苷酸(ODN_S)。测定了它们在S180肿瘤细胞中对VEGF表达的抑制作用,还测定了它们在鼠角膜肿瘤模型上抑制由肿瘤引起的新生血管形成和肿瘤的生长。结果表明,部分硫代的或发夹结构修饰的ODNs均可有效抑制VEGF的表达、肿瘤的生长和新生血管的形成。线性的ODN效果不好。两种作用于不同位点的ODN联合使用可得到最好的效果。因此反义VEGF寡聚脱氧核苷酸有望成为抗肿瘤和抗新生血管形成的新型药物。     

柱色谱法对大豆低聚糖浆脱色效果的研究

研究了离子交换树脂和大孔吸附树脂对大豆低聚糖浆的脱色效果。比较了不同树脂处理大豆低聚糖浆的脱色效果,并选择D392作为脱色树脂,确定的适宜操作条件为:固体分质量分数为10.75%的低聚糖浆与树脂的体积比为11∶1,在常温脱色3~4 h,脱色率可达86%以上,较优解吸剂为w(NaOH)=10%水溶液和无水乙醇的混合溶液V[w(NaOH)=10%的水溶液]∶V(无水乙醇)=1∶1,解吸率达99.81%。     

番茄红素的层析分离

以V(正己烷)∶V(丙酮)=1 5∶1 0的混合液为展开剂的硅胶薄层层析结果表明番茄红素油树脂中主要含有5种组分,番茄红素的Rf值为0 83;以V(正己烷)∶V(丙酮)=1 5∶1 0的混合液为洗脱剂,番茄红素油树脂经过硅胶柱层析分离,可得到w(番茄红素)=18 9%的番茄红素晶体产品;紫外可见光分光光度计检测结果表明,从番茄红素油树脂中还分离出β 胡萝卜素、叶黄素和玉米黄质。     

荧光光谱在核酸分析中的应用

讨论了荧光光谱及探针技术在核酸的结构、功能以及定性定量研究中的应用,指出荧光光谱法具有快速、简便、灵敏度高、选择性好等优点。     

柱层析法分离纯化碳酸甘油酯

采用氧化铝柱层析法分离纯化碳酸甘油酯,根据溶剂极性选择甲醇和丙酮混合溶液为洗脱剂,探讨了溶剂组成配比、粗碳酸甘油酯样品与氧化铝重量比和温度对分离效果的影响。由实验得出较佳分离条件为:洗脱剂配比V甲醇︰V丙酮=1︰19,粗碳酸甘油酯样品与氧化铝重量比N=1︰10,常温,此时纯度大于95%的碳酸甘油酯的收率可达92.53%。     

金属螯合亲和层析介质及应用研究的进展

金属螯合亲和层析具有螯合介质制备简单，吸附容量大，选择性及通用性较好，易于再生，成本低等优点，是分离纯化蛋白质等生物工程产品最有效的技术之一。本文综述了金属螯合亲和层析介质及最新应用方面的进展。     

毛细管气相色谱柱的选择和使用过程优化

阐述了选择毛细管柱的四个因素：固定相、柱长、内径、膜厚。介绍了大口径毛细管柱的优点及其对填充柱的取代。提出了使用毛细管柱时应注意的事项。