One-step capillary isoelectric focusing for the separation of the recombinant human immunodeficiency virus envelope glycoprotein glycoforms.
One-step capillary isoelectric focusing for the separation of the recombinant human immunodeficiency virus envelope glycoprotein glycoforms.
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一步毛细管等电聚焦用于分离重组人免疫缺陷病毒包膜糖蛋白糖型。
DOI：
10.1016/s0021-9673(99)01045-6
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
D. Ferrier
中科院分区：
文献类型：
--
作者：
N. Tran;M. Taverna;M. Chevalier;D. Ferrier
研究了一步毛细管等电聚焦法快速分离重组人免疫缺陷病毒（HIV）包膜糖蛋白（rgp 160 sMN/LAI）糖型的方法。在聚乙烯醇（PVA）涂层毛细管中使用窄和宽pH范围的两性电解质的混合物进行分离。已证明，蔗糖和3-（环己基氨基）-1-丙磺酸的组合是避免在接近其pI的pH下发生的蛋白质沉淀的最有效的添加剂。尽管校准曲线[等电点（pI）vs.迁移时间]显示非线性关系，但在短pI范围内可产生足够的线性，从而显示rgp 160 s MN/LAI不同糖型的酸性特征（pI为4.00 - 4.95）。通过比较聚丙烯酰胺和PVA涂层毛细管的性能进行的重现性评价表明，使用PVA毛细管进行的日内（0.5 - 1.9%）和日间（1.6 - 7.6%）测量的RSD值较低。此外，PVA毛细管的长期稳定性通过测量蛋白质标记物在长期使用中的迁移时间的变化来证明。最后，该方法能够区分两种接近的糖蛋白的糖型模式，例如病毒HIV-1的两个亚群的rgp 160。
One-step capillary isoelectric focusing was investigated as a rapid method to resolve the glycoforms of the heterogeneous recombinant human immunodeficiency virus (HIV) envelope glycoprotein (rgp 160sMN/LAI). The separation was performed in a poly(vinyl alcohol) (PVA) coated capillary using a mixture of ampholyte of narrow and wide pH range. A combination of saccaharose and 3-(cyclohexylamino)-1-propanesulfonic acid was shown to be the most efficient additive to avoid protein precipitation which occurs at a pH close to its pI. Although the calibration curve [isoelectric point (pI) vs. migration times] showed a non-linear relationship, an adequate linearity could be yielded for short pI ranges permitting to exhibit the acidic character of the different glycoforms of the rgp 160s MN/LAI (pI from 4.00 to 4.95). Reproducibility evaluated by comparing the performance of a polyacrylamide and a PVA coated capillary showed that low RSD values were obtained for intra-day (0.5 to 1.9%) and inter-day (1.6 to 7.6%) measurements using the PVA capillary. Moreover, the long term stability of the PVA capillary was demonstrated by measuring the variation of migration times of the protein markers for a long period of use. Finally, this method was able to differentiate the glycoform pattern of two close glycoproteins such as the rgp 160 of two sub-populations of the virus HIV-1.
毛细管区带电泳、毛细管等电聚焦和十二烷基硫酸钠毛细管电泳检查用于分析重组组织纤溶酶原激活剂。
DOI：
10.1016/0021-9673(96)00513-4
发表时间：
1996
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Thorne,JM;Goetzinger,WK;Chen,AB;Moorhouse,KG;Karger,BL
通讯作者：
Karger,BL