Isolation and characterization of monoclonal antibodies to Shiga-like toxin II of enterohemorrhagic Escherichia coli and use of the monoclonal antibodies in a colony enzyme-linked immunosorbent assay

Abstract

The major obstacle in large-scale epidemiological investigations of the incidence of Shiga-like toxin (SLT)-producing Escherichia coli in diarrheal stools is the lack of a rapid, specific test to detect toxin. Enterohemorrhagic E. coli produces elevated levels of SLT-I, SLT-ll, or both cytotoxins (also called Verotoxins). SLT-I but not SLT-II can be neutralized by antiserum to purified Shiga toxin and by monoclonal antibodies to the B subunit of SLT-I. In this study, monoclonal antibodies were generated against a crude preparation of SLT-II produced by an E. coli K-12 strain lysogenized with the 933W toxin-converting phage of enterohemorrhagic E. coli 933. Hybridoma culture supernatants were screened for anti-SLT-II antibodies by a cytotoxicity neutralization assay and by an enzyme-linked immunosorbent assay (ELISA). Of 53 ELISA-positive lines, 5 were capable of neutralizing the cytotoxicity of SLT-ll but not of SLT-I, Shiga toxin, or a variant of SLT-II produced by E. colu that causes edema disease of swine. All five monoclonal antibodies * Corresponding author.

Studies of Escherichia coli Cultured on RainbowTM Agar O157 with Particular Reference to Enterohaemorrhagic Escherichia coli (EHEC)

“…They were tested for their ability to produce SLT by a cell assay using vero cells (15). All strains were also tested by the ELISA technique described by Acheson (20), were used to confirm the presence of toxin(s). In many cases, the presence of the genes coding for the SLT were tested by a polymerase chain reaction (PCR) technique currently being developed in our laboratory.…”

Reliability of CHROMagar ® O157 for the detection of enterohaemorrhagic Escherichia coli (EHEC) O157 but not EHEC belonging to other serogroups

“…They were tested for their ability to produce SLT by the cell assay using vero cells (Konowalchuk et al 1977). All strains were also tested by the ELISA technique described by Acheson et al (1990) (Perera et al 1988), were used to confirm presence of the toxin(s). In many cases the presence of the genes coding for the SLT was tested for by the polymerase chain reaction technique, currently being developed in our laboratory.…”

Characterization of Monoclonal Immunoglobulin A and G Against Shiga Toxin Binding Subunits Produced by Intranasal Immunization

“…However, passive antibody therapy may hold promise. Murine monoclonal antibodies (mAbs) against Stx have been reported to neutralize the activity of Stx1 and ⁄ or Stx2 in vitro [16][17][18][19][20][21]. Stx1-neutralizing mAbs will have potential clinical utility for the prevention and treatment of HUS caused by Stx1-producing EHEC.…”