消除聚山梨酯80干扰后用考马斯亮蓝法进行蛋白质定量的快速方法。
Rapid method for protein quantitation by Bradford assay after elimination of the interference of polysorbate 80.
作者信息
Cheng Yongfeng, Wei Haiming, Sun Rui, Tian Zhigang, Zheng Xiaodong
机构信息
Institute of Immunology and CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Life Sciences and Medical Center, University of Science and Technology of China, Hefei, Anhui 230027, China.
Institute of Immunology and CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Life Sciences and Medical Center, University of Science and Technology of China, Hefei, Anhui 230027, China; Hefei National Laboratory for Physical Sciences at Microscale, Hefei, Anhui 230027, China.
出版信息
Anal Biochem. 2016 Feb 1;494:37-9. doi: 10.1016/j.ab.2015.10.013. Epub 2015 Nov 9.
Bradford assay is one of the most common methods for measuring protein concentrations. However, some pharmaceutical excipients, such as detergents, interfere with Bradford assay even at low concentrations. Protein precipitation can be used to overcome sample incompatibility with protein quantitation. But the rate of protein recovery caused by acetone precipitation is only about 70%. In this study, we found that sucrose not only could increase the rate of protein recovery after 1 h acetone precipitation, but also did not interfere with Bradford assay. So we developed a method for rapid protein quantitation in protein drugs even if they contained interfering substances.
考马斯亮蓝法是测量蛋白质浓度最常用的方法之一。然而，一些药用辅料，如洗涤剂，即使在低浓度下也会干扰考马斯亮蓝法。蛋白质沉淀可用于克服样品与蛋白质定量分析的不相容性。但丙酮沉淀导致的蛋白质回收率仅约为70%。在本研究中，我们发现蔗糖不仅可以提高丙酮沉淀1小时后的蛋白质回收率，而且不会干扰考马斯亮蓝法。因此，我们开发了一种即使蛋白质药物含有干扰物质也能快速进行蛋白质定量分析的方法。
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