Methodology for assaying recombinant interleukin-2 associated with liposomes by combined gel exclusion chromatography and fluorescence
Methodology for assaying recombinant interleukin-2 associated with liposomes by combined gel exclusion chromatography and fluorescence
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DOI：
10.1016/s1570-0232(02)00530-5
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发表时间：
2003-01-05
影响因子：
3
通讯作者：
Barratt, G
中科院分区：
文献类型：
--
作者：
Pellequer, Y;Ollivon, M;Barratt, G
建立了一种基于荧光和凝胶排斥色谱（GEC）的简单方法来测定与囊泡相关的重组白介素-2 （il -2）。采用Sephadex G75色谱柱分离脂质体与未包裹的il -2。采用荧光光散射法监测il -2脂质体的洗脱。在测定前用辛基葡萄糖苷（OG）溶解囊泡是必要的，以避免光散射的干扰。这种方法可以自动产生一个在线系统，可以分离，溶解和定量脂质体样品中的rIL-2。它可以扩展到与囊泡相关的任何蛋白质，只要前者可以被荧光检测到。(C) 2002 Elsevier Science B.V.版权所有
A simple methodology based on fluorescence and gel exclusion chromatography (GEC) has been developed to assay recombinant Interleukin-2 (rIL-2) associated with vesicles. A Sephadex G75 column was used to separate the liposomes from non-entrapped rIL-2. The elution of the rIL-2 liposomes was monitored by coupling fluorescent and light scattering detection. The solubilisation of the vesicles with octylglucoside (OG) before the assay was necessary to avoid interference from light scattering. This methodology can be automated to yield an on-line system that can separate, solubilise and quantify rIL-2 in liposome samples. It can be extended to any protein associated with vesicles provided that the former can be detected by fluorescence. (C) 2002 Elsevier Science B.V. All rights reserved.