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重组人干扰素—γ的凝胶色谱分离及其变性剂对分离结果的影响 总被引:1,自引:0,他引:1
使用十二烷基硫酸钠,盐酸胍和尿素三种不同的变性剂溶解基因重组人干扰素-Gamma(r-IFN-γ)包含体,然后进行凝胶色谱分离。使用活性,电泳和高效疏水作用色谱三种方法,比较了三种变性剂对分离结果的影响,结果为:尿素溶解包含体后r-IFN-γ的分子量与杂质相差较大,盐酸胍溶解液经分离后r-IFN-γ比活最高,SDS对r-IFN-γ的构象影响较大。 相似文献
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环糊精—十二烷基硫酸钠胶束电动色谱分离光学异构体的模型及色谱… 总被引:1,自引:0,他引:1
建立了环糊精-十二烷基硫酸钠胶电动色谱中可电离及电中性-光学异构体的分离模型。通过计算机模拟预测了多种操作因素对手性分离可能产生的影响,并通过对丹磺酰化氨基酸琢十八甲基炔诺酮旋光导松体的分离对理论预测进行了验证。 相似文献
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利用国产大孔硅胶作基质合成了疏水填料。按照高效疏水作用色谱法,采用梯度洗脱方式分离了6种标准蛋白及唾液中α-淀粉酶和基因工程生产的γ-干扰素。柱子不可逆吸附小、被试验的α-淀粉酶和溶菌酶活性几乎定量被回收。应用合成的色谱填料研究了洗脱剂中盐浓度和温度对蛋白质保留行为的影响,论证了合成填料的色谱属性。 相似文献
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分离蛋白质的高效离子交换色谱填料的研究(Ⅳ):强阴离子交换色… 总被引:1,自引:0,他引:1
常建华 《高等学校化学学报》1995,16(11):1737-1739
含环氧基的硅与二乙胺或聚乙烯亚胺反应,其产物再与碘甲烷或氯化苄反应,得到含季铵离子的强阴离子交换色谱填料,该埴料可用于蛋白质和生物工程产品(如基因重组人干扰素γ,粒细胞-巨噬细胞集落刺激因子和人绒毛膜促性腺素等)的分离纯化,其中,环氧基硅胶与二乙胺加成后再用氯化季铵化的产物是较好的强阴离子交换色谱填料。 相似文献
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毛细管离子交换电色谱的分离行为 总被引:7,自引:0,他引:7
在离子交换毛细管色谱柱上实施电色谱,并对其分离行为进行了研究,采用75μm(i.d.)20cm的毛细管强阳离子交换柱(3μm)以NaH2PO4-H3PO4缓冲液为淋洗剂,紫外柱上检测(214nm)考察了流动相的pH值,有机改性剂及分离电压等因素对分离的影响,研究表明,不同的pH溶质的流出次序发生改变,随着有机改性剂含量增加,溶质的保留时间减小,而电渗流却增大,同时,对分离的柱效和方法的重现性进行了 相似文献
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肿瘤坏死因子α和γ-干扰素对白血病细胞作用的31PNMR分析 总被引:3,自引:0,他引:3
用^31P核磁共振观察了肿瘤坏死因子α(TNF-α)和γ-干扰素(IFN-γ)联合对人淋巴瘤白血病细胞系Molt-4作用后含磷化合物的变化;Molt-4细胞系的^31P谱主要由三磷酸腺苷(ATP)、无机磷(Pi)、磷酸单酯等的共振峰组成;^31P谱显示γ-干扰素、肿瘤坏死因子α及γ-干扰素和肿瘤坏死因子α联合对Molt-4细胞作用后,三磷酸腺式苷的β位磷的共振峰与无机磷的峰高比值(hATP/hPi)不同程度地降低;同时根据Pi的化学位移对Molt-4细胞胞内pH的变化进行了监测。 相似文献
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化学合成虎纹捕鸟蛛毒素-Ⅰ的色谱行为及分离纯化 总被引:4,自引:0,他引:4
利用反相高效液相色谱 (RP HPLC)和离子交换色谱 (IEC)比较研究了固相化学合成的多肽类神经毒素虎纹捕鸟蛛毒素 Ⅰ (HWTX Ⅰ )与其对应的天然产物的色谱行为差异及分离纯化方法。发现在完全相同的实验条件下 ,复性处理后的合成HWTX Ⅰ的谱峰有别于还原变性后又复性的天然HWTX Ⅰ的谱峰 ,主要表现为扩散度较大 ,对称性较差 ,说明合成产物存在更大分子构象的不均一性。推测除了在合成HWTX Ⅰ复性过程中有少数分子发生二硫键错配外 ,还有部分分子在固相合成中发生了消旋作用 ,需要交替采取多种色谱法才能完全分离纯化复性后的合成多肽产物。 相似文献
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BAI Quan KONG Yu DONG Cuihua & GENG Xindu Institute of Modern Separation Science Shaanxi Key Laboratory of Modern Separation Science Northwest University Xi''''an China 《中国科学B辑(英文版)》2005,48(Z1)
The refolding of the reduced-denatured insulin from bovine pancreas was investigated with the size exclusion chromatography (SEC). It was shown that the reduced-denatured insulin originally denatured with 7.0 mol·L-1 guanidine hydrochloride (GuHCI) or 8.0 mol·L-1 urea could not be refolded with a non-oxidized mobile phase. Although the oxidized and reduced glutathione (GSSG and GSH) were employed in the oxidized mobile phase, the reduced-denatured insulin still could not be renatured. However, in the presence of 2.0 mol·L-1 urea in the oxidized mobile phase employed, the reduced-denatured insulin can be refolded with SEC, and the aggregation of denatured insulin can be diminished by urea. In addition, the disul-fide exchange of reduced-denatured insulin also can be accelerated with GSSG/GSH in the oxidized mobile phase. The three disulfide bridges of insulin were formed correctly and the reduced-unfolded insulin can be renatured completely. The results were further tested with re-versed-phase liquid chromatography (RPLC) and hydrophobic interaction chromatography (HIC). 相似文献
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A commercial Chromolith C18 column and two new stationary phases with mixed ligands bonded on the Kromasil silica gel support, SG-MIX and SG-Chol, were characterized using simple tests based on the retention of non-polar, basic and acidic compounds. Polar and methylene selectivity tests in acetonitrile-water and methanol-water mobile phases revealed lower hydrophobicities of the SG-MIX and SG-Chol columns in comparison to the Chromolith column. The columns were further characterized using new test criteria - gradient oligomer capacity and isomeric selectivity and peak symmetry of naphthalene di-sulphonic acids in aqueous mobile phases. The cholesterolic column shows greater gradient oligomer selectivity for the separation of oligoethylene glycol samples than the SG-MIX and the Chromolith columns. Increased retention and peak tailing, but decreased isomeric selectivity for naphthalene-di-sulphonic acids was observed with the SG-MIX column, because of interactions with various polar bonded groups. 相似文献
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The refolding of the reduced-denatured insulin from bovine pancreas was investigated with the size exclusion chromatography (SEC). It was shown that the reduced-denatured insulin originally denatured with 7.0 mol L?1 guanidine hydrochloride (GuHCI) or 8.0 mol L?1 urea could not be refolded with a non-oxidized mobile phase. Although the oxidized and reduced glutathione (GSSG and GSH) were employed in the oxidized mobile phase, the reduced-denatured insulin still could not be renatured. However, in the presence of 2.0 mol Lt-1 urea in the oxidized mobile phase employed, the reduced-denatured insulin can be refolded with SEC, and the aggregation of denatured insulin can be diminished by urea. In addition, the disulfide exchange of reduced-denatured insulin also can be accelerated with GSSG/GSH in the oxidized mobile phase. The three disulfide bridges of insulin were formed correctly and the reduced-unfolded insulin can be renatured completely. The results were further tested with reversed-phase liquid chromatography (RPLC) and hydrophobic interaction chromatography (HIC). 相似文献
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The impact of column inner diameter on chromatographic performance in temperature gradient liquid chromatography has been investigated in the present study. Columns with inner diameters of 0.32, 0.53, 3.2 and 4.6 mm were compared with respect to retention and efficiency characteristics using temperature gradients from 30 to 90 degrees C with temperature ramps of 1, 5, 10 and 20 degrees C min(-1). The columns were all of 15 cm length and were packed with 3 microm Hypersil ODS particles. Alkylbenzenes served as model compounds, and the mobile phase consisted of acetonitrile-water (50:50, v/v). The study revealed that the column ID is not a critical limiting factor when performing temperature programming in LC, at least for columns narrower than 4.6 mm inner diameter in the temperature interval 30-90 degrees C. The retention times for all components on all columns were highly comparable, with similar peak profiles without any signs of peak splitting. The use of mobile phase pre-heating when using the larger bore columns was avoided by starting the temperature gradients close to ambient. However, the relative apparent efficiency was inversely proportional to column inner diameter, making the capillary columns generally more functional towards temperature gradients than the larger bore columns with respect to chromatographic efficiency. In addition, the capillary columns possessed higher robustness towards temperature programming than the conventional columns. 相似文献
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The refolding of the reduced/denatured insulin from bovine pancreas as the model protein was investigated with weak anion exchange chromatography(WAX) coupled with MALDI-TOF MS.The results indicated that the disulfide bonds almost cannot be formed correctly with the common mobile phase by WAX.However,with the urea gradient elution and in the presence of GSSG/ Cyst as the ratio 1:6 in the mobile phase employed,the disulfide exchange of reduced/denatured insulin can be accelerated resulting in forming the ... 相似文献
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A new mathematical treatment concerning the gradient elution in reversed-phase liquid chromatography when the volume fraction psi of an organic modifier in the water-organic mobile phase varies linearly with time is presented. The experimental ln k versus psi curve, where k is the retention factor under isocratic conditions in a binary mobile phase, is subdivided into a finite number of linear portions and the solute gradient retention time tR is calculated by means of an analytical expression arising from the fundamental equation of gradient elution. The validity of the proposed analytical expression and the methodology followed for the calculation of tR was tested using eight catechol-related solutes with mobile phases modified by methanol or acetonitrile. It was found that in all cases the accuracy of the predicted gradient retention times is very satisfactory because it is the same with the accuracy of the retention times predicted under isocratic conditions. Finally, the above method for estimating gradient retention times was used in an optimisation algorithm, which determines the best variation pattern of psi that leads to the optimum separation of a mixture of solutes at different values of the total elution time. 相似文献