Key Laboratory of Bio-Resources and Eco-Environment Ministry of Education/Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu, P R China.
J Sep Sci. 2011 Nov;34(21):3092-8. doi: 10.1002/jssc.201100235. Epub 2011 Oct 12.
α-Momorcharin (α-MMC), a type I ribosome-inactivating protein (RIP), has shown therapeutic potential such as anti-tumor and anti-viral agent. Traditional process of α-MMC purification from bitter melon seeds was time consuming and low efficient. To take this challenge, we made an affinity matrix by coupling the monoclonal antibody (McAb) with Sepharose 4B. Using this attractive strategy, 196 mg of α-MMC was obtained from 100 g of bitter melon seeds as the starting material. The yield of the protein was 2.7%. The homogeneity and properties of the protein were assessed by SDS-PAGE, acidic PAGE, RP-HPLC and N-terminal sequence as well as Western blot. Purified α-MMC showed remarkable inhibition to the melanoma cell line JAR and EMT-62058. In addition, it also displayed obvious inhibition on hepatitis B virus (HBV). This work provided a simple, rapid and efficient approach for α-MMC purification from Momordica charantia.
α-苦瓜素（α-MMC），一种 I 型核糖体失活蛋白（RIP），具有抗肿瘤和抗病毒等治疗潜力。从苦瓜种子中纯化α-MMC 的传统方法既耗时又低效。为了应对这一挑战，我们通过将单克隆抗体（McAb）与 Sepharose 4B 偶联来制备亲和基质。利用这一有吸引力的策略，从 100g 苦瓜种子起始材料中获得了 196mg 的α-MMC。该蛋白的产率为 2.7%。通过 SDS-PAGE、酸性 PAGE、RP-HPLC 和 N 端序列以及 Western blot 评估了蛋白质的均一性和性质。纯化的α-MMC 对黑素瘤细胞系 JAR 和 EMT-62058 表现出显著的抑制作用。此外，它还对乙型肝炎病毒（HBV）表现出明显的抑制作用。这项工作为从苦瓜中纯化α-MMC 提供了一种简单、快速、高效的方法。
