Production and characterisation of monoclonal antibodies against pyelonephritis-associated P-pili of Escherichia coli.
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.
Miyata
H
H
Department of Paediatrics, Kinki University School of Medicine, Osaka, Japan.
Kataoka
S
S
Moriguchi
N
N
Yamamoto
T
T
Michibata
I
I
Kobayashi
T
T
Maki
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology* / Q000302:isolation & purification / Q000648:ultrastructure; D004927:Escherichia coli Infections / Q000382:microbiology; D036781:Fimbriae Proteins; D005455:Fluorescent Antibody Technique; D006386:Hemagglutination Tests; D007075:Immunoglobulin M / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D018933:Pili, Sex / Q000737:chemistry / Q000276:immunology* / Q000648:ultrastructure; D011704:Pyelonephritis / Q000382:microbiology*
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D007075:Immunoglobulin M; D036781:Fimbriae Proteins