Preparation and Primary Application of Monoclonal Antibodies against a Novel Ribosome-inactivating Protein Moschatin from Pumpkin Seeds

查看全文 作　　者：Heng-ChuanXIA Wei-GuoHU Xin-XiuYANG FengLI Zu-[1]ChuanZHANG

机构地区：[1]KeyLaboratoryofProteomics,InstituteofBiochemistryandCellBiology,ShanghaiInstitutesforBiologicalSciences,theChineseAcademyofSciences,Shanghai200031,China

出　　处：《Acta Biochimica et Biophysica Sinica》索引2004年第36卷第2期,共6页

摘　　要：Plant ribosome-inactivating proteins (RIPs) have multiple biological functions, and have beenwidely used in the studies on biomedical and agronomic applications. Moschatin is a novel single-chain RIPrecently purified from pumpkin seeds, and it has been successfully applied to construct the immunotoxin thatcan selectively kill the cultured human melanoma cells. Six stable strains of hybridomas (2H8, 4A8, 5B6,6F8, 4H10 and 6C2) that can secrete high specific monoclonal antibodies against Moschatin have beensuccessfully prepared using hybridoma technique. The isotypes of these monoclonal antibodies areIgG1, IgG1, IgG1, IgG1, IgG2a and IgM. Their affinity constants were determined to be 1.42×108, 2.71×108,8.72×07, 2.06×l08, 1.36×108 and 1.51×108 M-1 in a sequent order, measured by non-competitive ELISA.The monoclonal antibody 4A8 has been used to detect Moschatin in Western blot. An immunoaffinity gel,which consisted ofa monoclonal antibody 4H 10 and Sepharose 4B, was prepared and used to purify Moschatinfrom pumpkin seeds crude extract. 关键词：南瓜种子核糖体钝化蛋白单克隆抗体免疫毒素黑素瘤细胞人类纯化

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Preparation and Primary Application of Monoclonal Antibodies against a Novel Ribosome-inactivating Protein Moschatin from Pumpkin Seeds

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