A monoclonal antibody for the specific diagnosis of plague

Abstract

A stable mouse-cell hybridoma was obtained that secretes an IgA monoclonal antibody reactive with the fraction 1 (F1) envelope antigen of Yersinia pestis. Titres of the antibody typically ranged from 1:32 768 to 1:65 536 in mouse ascitic fluids.The monoclonal antibody formed a line of precipitation when run against F1 antigen in Ouchterlony gel diffusion tests. In tests of 235 strains of Y. pestis, lines of identity occurred between the precipitates formed with a solution of purified F1 antigen and the F1 antigen produced by the plague strains. No precipitates formed for 65 strains that were incapable of elaborating F1 antigen. Specificity of the monoclonal antibody for strains of Y. pestis producing F1 was also indicated by negative results for 50 yersinia strains other than Y. pestis tested by an ELISA that used the antibody to capture antigen.Experiments to determine the shelf-life of the antibody were conducted over 3-4 years. When the monoclonal antibody was freeze-dried in vials, titre was retained for three years when the vials were stored at -70 degrees C but only for two months when they were stored at ambient temperatures. When the antibody was freeze-dried in wells of ELISA plates, sensitivity of the plates for capture of F1 antigen was preserved for four years when the plates were stored at -70 degrees C compared with two weeks for plates stored at room temperature. When a solution of the antibody was sealed in wells of ELISA plates and refrigerated at 4 degrees C, reactivity of the antibody and sensitivity of the plates were retained for a year.Alternatives for the application of this monoclonal antibody in ELISA and other plague diagnostic procedures are discussed.