Immunoaffinity purification of a-momorcharin from bitter melon seeds (Momordica charantia)

Abstract

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, RPHPLC 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. © 2011 WILEY-VCH Verlag GmbH &Co. KGaA, Weinheim.