Use of a Resin Bound Monoclonal Antibody in the Purification of the Various Components in Anthrax Vaccines

摘要

The results of the research supported by this contract have been recorded in eight papers. The papers can be divided into two equal groups. The first describes the purification techniques performed to independently isolate the three exotoxins of Bacillus anthracis: protective antigen (PA), lethal factor (LF) and edema factor (EF). The sequential purification technique of the three exotoxins is described in an additional paper. These papers delineate a good technique for isolating all of the exotoxins individually using columns prepared with their own specific monoclonal antibody. An alternative method is to purify the crude fermentation bacterial broth by removing PA and LF and allowing the residue to serve as EF. This method is almost as effective as preparing EF alone from its own specific monoclonal antibody columns. The specific points uncovered by our research are herein summarized in paragraphs corresponding to the paper in which they were or will be published. The main topics are: Studies on the specific exotoxins of Bacillus anthracis. Purification of lethal factor. Protective antigen. Sequential purification of Bacillus anthracis exotoxins by immunosorbent chromatography. Purification of EF by immunosorbent chromatography. Studies performed on immunochromatography techniques. The stability of antibody attachment in immunosorbent chromatography. Effect of borohydride reduction on antibodies. Evaluation of activation methods with cellulose beads for the immunosorbent purification of immunoglobulins. Immobilization of antibody on cellulose bead derivatives.