抗大肠杆菌肾盂肾炎相关P菌毛单克隆抗体的制备与特性分析

Production and characterisation of monoclonal antibodies against pyelonephritis-associated P-pili of Escherichia coli.

作者信息

Miyata H, Kataoka S, Moriguchi N, Yamamoto T, Michibata I, Kobayashi T, Maki S

机构信息

Department of Paediatrics, Kinki University School of Medicine, Osaka, Japan.

出版信息

Pediatr Nephrol. 1994 Jun;8(3):270-4. doi: 10.1007/BF00866329.

DOI:10.1007/BF00866329

PMID:7917847

Abstract

Pyelonephritis-associated P-pili (PAP) of Escherichia coli O6,H(-),K1(-),F12,haemolysin(-) were purified by salt precipitation and affinity chromatography using Synsorb P1. Purified PAP showed a single band with a molecular weight of 18 kDa by electrophoretic analysis. A monoclonal antibody (mAb) was produced by fusion of the PAI myeloma cell line with splenic lymphocytes from BALB/c mice immunised with the purified PAP. The mAb was of IgM class with kappa light chains and reacted with a 18-kDa moeity of the salt precipitate; the epitope was present near the apical part of the pilus filaments. The mAb reacted with PAP in both immunofluorescence and haemagglutination tests when 108 strains isolated from urine samples were tested; the two tests were in agreement for 202 of 204 strains isolated from faecal samples.

摘要

通过盐沉淀和使用Synsorb P1的亲和层析法纯化了大肠杆菌O6、H(-)、K1(-)、F12、溶血素(-)的肾盂肾炎相关P菌毛（PAP）。电泳分析显示纯化的PAP呈现一条分子量为18 kDa的条带。通过将PAI骨髓瘤细胞系与用纯化的PAP免疫的BALB/c小鼠的脾淋巴细胞融合，制备了单克隆抗体（mAb）。该mAb为IgM类，具有κ轻链，与盐沉淀的18 kDa部分发生反应；表位存在于菌毛丝的顶端附近。当对从尿液样本中分离出的108株菌株进行测试时，该mAb在免疫荧光和血凝试验中均与PAP发生反应；从粪便样本中分离出的204株菌株中有202株在这两项试验中结果一致。