SEPARATION OF ANTIBODY-ANTIGEN COMPLEXES BY CAPILLARY ZONE ELECTROPHORESIS, ISOELECTRIC-FOCUSING AND HIGH-PERFORMANCE SIZE-EXCLUSION CHROMATOGRAPHY
SEPARATION OF ANTIBODY-ANTIGEN COMPLEXES BY CAPILLARY ZONE ELECTROPHORESIS, ISOELECTRIC-FOCUSING AND HIGH-PERFORMANCE SIZE-EXCLUSION CHROMATOGRAPHY
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DOI：
10.1016/s0021-9673(01)95371-3
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发表时间：
1991-02-08
期刊：
影响因子：
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通讯作者：
SITTAMPALAM, GS
中科院分区：
文献类型：
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作者：
NIELSEN, RG;RICKARD, EC;SITTAMPALAM, GS
自由溶液毛细管区带电泳（CZE）分离抗体-抗原复合物已得到证实。抗原、对抗原特异的单克隆抗体和复合物被很好地分离。采用柱上紫外检测，在不到10 min的时间内实现了整个分离。通过聚丙烯酰胺凝胶上的等电聚焦（IEF）实验，分别估计了这三个物种的pI值。通过CZE测量的相对迁移时间和pI值之间存在合理的良好一致性。抗体-抗原混合物的IEF和高性能尺寸排阻色谱证实了CZE观察到的复合物的形成。这项研究表明，实用程序的CZE作为一种新的和互补的技术表征的抗体-抗原复合物。
The separation of antibody-antigen complexes by free-solution capillary zone electrophoresis (CZE) has been demonstrated. The antigen, a monoclonal antibody specific for the antigen, and the complex were well resolved. The entire separation was achieved in less than 10 min using on-column UV detection. The pI values for the three species were estimated separately by isoelectric focusing (IEF) experiments on polyacrylamide gels. Reasonably good agreement was found between the relative migration times measured by CZE and the pI values. Both IEF and high-performance size-exclusion chromatography of the antibody-antigen mixtures confirmed the formation of the complexes observed by CZE. This study demonstrates the utility of CZE as a new and complementary technique for the characterization of antibody-antigen complexes.