针对黄疸出血型问号钩端螺旋体EDTA提取物的单克隆抗体的制备与特性研究

Production and characterization of monoclonal antibodies to the EDTA extract of Leptospira interrogans, serovar icterohaemorrhagiae.

作者信息

Leite L T, Resende M, de Souza W, Camargos E R, Koury M C

机构信息

Departmento de Microbiologia e Morfologia do Instituto de Ciencias Biolgicas, Universidade Federal de Minas Gerais, Belo Horizonte.

出版信息

Rev Soc Bras Med Trop. 1996 Sep-Oct;29(5):483-9. doi: 10.1590/s0037-86821996000500012.

DOI:10.1590/s0037-86821996000500012

PMID:8885673

Abstract

Monoclonal antibodies (MABs) were produced against an ethylenediaminetetraacetate (EDTA) extract of Leptospira interrogans serovar icterohaemorrhagiae being characterized by gel precipitation as IgM and IgG (IgG1 and IgG2b). The EDTA extract was detected as several bands by silver staining in SDS-PAGE. In the Western blot the bands around 20 KDa reacted with a monoclonal antibody, 47B4D6, and was oxidized by periodate and was not digested by pronase, suggesting that the determinant is of carbohydrate nature. Immunocytochemistry, using colloidal gold labeling, showed that an EDTA extract determinant recognized by monoclonal antibody 47B4D6, is localized under the outer envelope of serovar icterohaemorrhagiae. The MAB raised against the EDTA extract was not able to protect hamsters from lethal challenge with virulent homologous leptospires.

摘要

制备了针对问号钩端螺旋体黄疸出血型乙二胺四乙酸（EDTA）提取物的单克隆抗体（MABs），通过凝胶沉淀法鉴定其为IgM和IgG（IgG1和IgG2b）。在SDS-PAGE中，通过银染法检测到EDTA提取物呈现多条条带。在蛋白质印迹法中，约20 kDa处的条带与单克隆抗体47B4D6发生反应，可被高碘酸盐氧化且不被链霉蛋白酶消化，表明该决定簇具有碳水化合物性质。采用胶体金标记的免疫细胞化学显示，单克隆抗体47B4D6识别的EDTA提取物决定簇位于黄疸出血型血清型的外膜之下。针对EDTA提取物产生的单克隆抗体无法保护仓鼠免受同源强毒钩端螺旋体的致死性攻击。