抗体偶联葡萄糖氧化酶对人内皮细胞的特异性杀伤作用

Specific killing of human endothelial cells by antibody-conjugated glucose oxidase.

作者信息

Muzykantov V R, Sakharov D V, Sinitsyn V V, Domogatsky S P, Goncharov N V, Danilov S M

机构信息

Institute of Experimental Cardiology, National Cardiology Research Center of Academy of Medical Sciences of USSR, Moscow.

出版信息

Anal Biochem. 1988 Mar;169(2):383-9. doi: 10.1016/0003-2697(88)90300-4.

DOI:10.1016/0003-2697(88)90300-4

PMID:3260084

Abstract

Conjugates of antibody with glucose oxidase obtained via the carbohydrate moiety of the enzyme via oxidation with periodate are suggested as a tool for selective killing of the target cell. The conjugate was separated from uncoupled enzyme by repeated precipitation with ammonium sulfate (42% saturation). Purity of the conjugate was estimated by gel filtration on Toya Pearl TP-65. The glucose oxidase conjugated with rabbit antibody against mouse IgG bound specifically to plastic-adsorbed mouse immunoglobulins and to cultured human endothelial cells pretreated with mouse anti-endothelial antiserum. Glucose oxidase targeted to the cells generates hydrogen peroxide in the presence of glucose. This hydrogen peroxide killed the endothelial cells even in the absence of a halide-peroxidase system and in the presence of catalase. Features of the conjugate (specificity, effective cytotoxicity, high stability) make it suitable for prospective in vivo application and for immunoselective segregation of heterogeneous cell populations.

摘要

通过高碘酸盐氧化，经由葡萄糖氧化酶的碳水化合物部分获得的抗体与葡萄糖氧化酶的结合物被认为是一种选择性杀伤靶细胞的工具。通过用硫酸铵（饱和度42%）反复沉淀，将结合物与未偶联的酶分离。结合物的纯度通过在Toya Pearl TP - 65上进行凝胶过滤来估计。与抗小鼠IgG的兔抗体偶联的葡萄糖氧化酶特异性结合到塑料吸附的小鼠免疫球蛋白以及用小鼠抗内皮抗血清预处理的培养人内皮细胞上。靶向细胞的葡萄糖氧化酶在葡萄糖存在下产生过氧化氢。即使在没有卤化物过氧化物酶系统且存在过氧化氢酶的情况下，这种过氧化氢也能杀死内皮细胞。结合物的特性（特异性、有效的细胞毒性、高稳定性）使其适用于未来的体内应用以及异质细胞群体的免疫选择性分离。