Wardale R J, Maciewicz R A, Etherington D J
Biosci Rep. 1986 Jul;6(7):639-46. doi: 10.1007/BF01114758.
Three stable hybridoma cell lines (AF8, BC11, CE2) have been produced that secrete antibodies specific for cathepsin B. These have been characterized by ELISA, SDS-PAGE immunostaining, immunoprecipitation and immunofluorescent staining. CE2 immunoprecipitated native cathepsin B with retention of enzymic activity, but failed to cross-react with the alkali-denatured enzyme. BC11 bound only to the denatured form of cathepsin B and AF8 cross-reacted with both native and denatured cathepsin B. However, unlike CE2-immunoprecipitated enzyme, activity could be detected only after dissociation of the antigen-AF8 antibody complex. No cross reaction was found with any lysosomal protein including the cysteine proteinases, cathepsins H and L.
已产生三种稳定的杂交瘤细胞系（AF8、BC11、CE2），它们分泌对组织蛋白酶B具有特异性的抗体。这些细胞系已通过酶联免疫吸附测定（ELISA）、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）免疫染色、免疫沉淀和免疫荧光染色进行了表征。CE2免疫沉淀天然组织蛋白酶B并保留酶活性，但与碱变性酶无交叉反应。BC11仅与组织蛋白酶B的变性形式结合，而AF8与天然和变性的组织蛋白酶B均发生交叉反应。然而，与CE2免疫沉淀的酶不同，只有在抗原-AF8抗体复合物解离后才能检测到活性。未发现与任何溶酶体蛋白（包括半胱氨酸蛋白酶组织蛋白酶H和L）发生交叉反应。
