Development of a monoclonal antibody capable of interacting with five serotypes of Staphylococcus aureus enterotoxin

Abstract

A monoclonal antibody capable of binding to determinants shared in common by staphylococcal enterotoxin serotypes A, B, Cl, D, and E was developed. To accomplish this, BALB/c mice were immunized by alternating injections of serotypes A and D to enrich for spleen lymphocytes programmed to produce antibody to possible common determinants. These cells, fused with mutant myeloma cells (P3-X-63Ag8.653) produced hybrids that formed monoclonal antibodies to either serotype A only or to both serotypes A and D. A cloned hybridoma from the latter group produced an antibody (subclass immunoglobulin Gl) which interacted with five serotypes. Highest affinity was to B and C1. An immunomatrix consisting of this antibody cross-linked to protein A-Sepharose CL-4B with dimethyl pimelimidate was capable of binding enterotoxin. Bound toxin was eluted with diethylamine. Because of its ability to interact with all five serotypes, the monoclonal antibody should prove useful in the development of a rapid method for screening foods for the presence of staphylococcal enterotoxin.

Murine monoclonal antibodies against staphylococcal enterotoxin B: production and characterization

“…Absorbances in each case (IgG~ or IgG2b) were markedly higher than in other subclasses (data not shown). The percentage of mAbs belonging to IgGzb subclass (42.85%) is much higher than those described by other authors for SEs [2,9,17] and most authors describe antibodies belonging only to lgG 1 subclass [6,10,14].…”

“…There are many reports on the development and use of monoclonal antibodies (mAbs) produced against SEs [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18]. mAbs to all of the identified enterotoxins have been prepared [2], either specific or cross-reactive.…”

“…mAbs to all of the identified enterotoxins have been prepared [2], either specific or cross-reactive. Meyer et al [14] prepared a monoclonal antibody which reacts with all of the identified enterotoxins, but it also reacts non-specifically with other proteins [1,19]. Edwin et al [7] produced mAbs which reacted specifically with SEA, SEB, SECt, SED and SEE.…”

Production, purification and identification of the staphylococcal enterotoxins

“…Several reports have described the production of mAbs against staphylococcal enterotoxins (Edwin et al 1984;Meyer et al 1984; Thompson et al 1984;Lin et al 1988). When a number of mAbs were tested in both RIA and ELISA systems (Thompson et al 1985(Thompson et al , 1986 it was found that single mAbs did not perform as well in RIA as polyclonal antibodies or a cocktail consisting of three or more mAbs.…”

Evaluation of an alternative extraction procedure for enterotoxin determination in dairy products

“…These procedures are time-consuming and may result in a low recovery of enterotoxins. Different modifications of enterotoxin purification have included chromatofocusing (Ende et al 1983;Fey et al 1984b;Lei et al 1988), development of dye ligand chromatography (Reynolds et al 1988) and the use of monoclonal antibody immunoaffinity chromatography (Meyer et al 1984;Lapeyre et al 1987;Shinagawa et al 1991). The dialysis concentration method against 30% polyethylene glycol (PM 20 000) has been used prior to detection of enterotoxin with the French reference method (Anon.…”