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Rapid method for protein quantitation by Bradford assay after elimination of the interference of polysorbate 80
Authors:Yongfeng Cheng  Haiming Wei  Rui Sun  Zhigang Tian  Xiaodong Zheng
Institution:1. 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;2. Hefei National Laboratory for Physical Sciences at Microscale, Hefei, Anhui 230027, China
Abstract: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.
Keywords:Sucrose  Protein precipitation  Bradford assay
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