Kurobe N, Inagaki T, Kato K
Department of Biochemistry, Aichi Prefectural Colony, Japan.
Clin Chim Acta. 1990 Dec 3;192(3):171-9. doi: 10.1016/0009-8981(90)90219-i.
A sensitive sandwich-type enzyme immunoassay for measurement of human Mn superoxide dismutase (Mn SOD) was developed using purified antibodies specific to Mn SOD. The antisera were raised in rabbits by injecting Mn SOD purified from human liver. The antibody IgG, purified by the use of Mn SOD-coupled Sepharose, showed a single band on the immunoblotting test with a crude liver extract. The assay system consisted of polystyrene balls with immobilized monospecific antibody F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The assay was highly sensitive and the minimum detection limit was 1 pg human Mn SOD/assay tube. Serum Mn SOD concentrations of healthy adults (77.5 +/- 18.0 ng/ml (1 SD), n = 120, 16-64 yr old) were not related to age or sex. Immunoreactive Mn SOD was detectable in most tissues examined except for erythrocytes. The concentrations of immunoreactive Mn SOD and Cu/Zn SOD in the cerebral cortex were not different among the patients with Alzheimer's disease, and the age matched and young patients without neurological disorders.
利用针对锰超氧化物歧化酶（Mn SOD）的纯化抗体，开发了一种用于检测人Mn SOD的灵敏夹心型酶免疫测定法。通过注射从人肝脏纯化的Mn SOD，在兔体内产生抗血清。使用与Mn SOD偶联的琼脂糖凝胶纯化的抗体IgG，在与粗制肝脏提取物的免疫印迹试验中显示出单一条带。该测定系统由固定有单特异性抗体F(ab')2片段的聚苯乙烯球和用来自大肠杆菌的β-D-半乳糖苷酶标记的相同抗体Fab'片段组成。该测定法高度灵敏，最低检测限为1 pg人Mn SOD/测定管。健康成年人（77.5±18.0 ng/ml（1个标准差），n = 120，16 - 64岁）的血清Mn SOD浓度与年龄或性别无关。除红细胞外，在大多数检测的组织中均可检测到免疫反应性Mn SOD。在患有阿尔茨海默病的患者、年龄匹配的无神经疾病的患者以及年轻患者中，大脑皮质中免疫反应性Mn SOD和铜/锌超氧化物歧化酶（Cu/Zn SOD）的浓度没有差异。
