Anthrax specific antibodies

Abstract

The present invention is directed to diagnostic tools and therapies using antibodies to Bacillus anthracis . Specifically, the present invention is directed to a B. anthracis -specific monoclonal antibody that binds to the EA1 antigen (corresponding to the eag gene) of the S-layer (surface layer) of spores. This monoclonal antibody may be used in a variety of applications, including to specifically detect and diagnose B. anthracis . Preferably, antibodies are monoclonal and bind to a surface protein, such as EA1 protein, on the spores of B. anthracis , and not to spores of either B. cereus or B. thuringiensis . Antibodies can be incorporated into detection kits using, for example, colloidal particle based lateral flow detection system. Such detection kits can distinguish anthrax spores from non-pathogenic varieties of spores. In addition, the invention is directed to B. anthracis EA1 antigen and pharmaceuticals such as vaccines that can be used as therapeutics and to develop improved antibodies and detection methods.

1 . A monoclonal antibody which is specifically reactive against  B. anthracis.    
     
     
         2 . The antibody of  claim 1  which is non-reactive against  B. cereus  or  B. thuringiensis.    
     
     
         3 . The antibody of  claim 1  which is an IgA, IgD, IgE, IgG or IgM. 
     
     
         4 . The antibody of  claim 1  which is reactive against a surface protein of  B. anthracis.    
     
     
         5 . The antibody of  claim 4  wherein the surface protein is an EA1 protein. 
     
     
         6 . The antibody of  claim 1  which binds to SEQ ID NO. 1. 
     
     
         7 . The antibody of  claim 1  which is specifically reactive against  B. anthracis  spores. 
     
     
         8 . The antibody of  claim 1  which is specifically reactive against  B. anthracis  vegetative cells. 
     
     
         9 . A hybridoma that produces the antibody of  claim 1 . 
     
     
         10 . The hybridoma of  claim 9  which is derived from an animal selected from the group consisting of cattle, chickens, goats, guinea pigs, horses, mice, pigs, primates, rabbits, rats and sheep. 
     
     
         11 . A hybridoma deposited with ATCC and accorded accession number PTA-2632. 
     
     
         12 . Antibody isolated from the hybridoma of  claim 11 . 
     
     
         13 . An isolated antibody, or reactive portion thereof, directed to the EA1 protein of  B. anthracis.    
     
     
         14 . The antibody or reactive portion thereof of  claim 13  which is a murine antibody; a rabbit antibody; a rat antibody; a genetically engineered antibody; a recombinant antibody; a humanized antibody; a polyclonal antibody or an affinity-purified antibody. 
     
     
         15 . The antibody or reactive portion thereof of  claim 13  which is an Fab or Fv fragment. 
     
     
         16 . A diagnostic kit comprising an antibody that is specifically reactive against spores or vegetative cells of  B. anthracis, B. thuringiensis  or  B. cereus.    
     
     
         17 . The diagnostic kit of  claim 16  which incorporates a colloidal particle based lateral flow detection system. 
     
     
         18 . The diagnostic kit of  claim 16  which incorporates a detection system selected from the group consisting of a carbon based lateral flow system; a fluorescent based assay system, a chemiluminescent system, an up converting phosphors system, a refractive indexed based detection system, a magnetic bead or latex bead system, and a micro array system. 
     
     
         19 . A diagnostic kit comprising an antibody that is specifically reactive against spores of  B. anthracis  and not  B. thuringiensis , and incorporates a colloidal particle based lateral flow detection system. 
     
     
         20 . A diagnostic kit comprising an antibody that is specifically reactive against spores of  B. thuringiensis  and not  B. anthracis , and incorporates a colloidal particle based lateral flow detection system. 
     
     
         21 . A method for producing a species-specific monoclonal antibody to one species of  Bacillus  comprising:
 immunizing a host with a preparation of said one species of  Bacillus;      boosting said host with another preparation of an antigenically similar, but not identical, species of  Bacillus;      boosting said host with said preparation of said one species;   fusing antibody-producing cells from the host with immortalized cells; and   selecting a hybridoma that produces the species-specific monoclonal antibody to said one species of  Bacillus.      
     
     
         22 . The method of  claim 21  wherein the one species of  Bacillus  is  B. cereus, B. thuringiensis  or  B. anthracis.    
     
     
         23 . The method of  claim 21  wherein the other antigenically similar species of  Bacillus  is  B. cereus, B. thuringiensis, B. anthracis  or combinations thereof. 
     
     
         24 . The method of  claim 21  wherein the one species of  Bacillus  is  B. anthracis  and the other antigenically similar species of  Bacillus  is  B. cereus  or  B. thuringiensis.    
     
     
         25 . The method of  claim 21  wherein the one species of  Bacillus  is  B. thuringiensis  and the other antigenically similar species of  Bacillus  is  B. cereus  or  B. anthracis.    
     
     
         26 . The method of  claim 21  wherein the preparation of said one species comprises spores, vegetative cells or combinations thereof. 
     
     
         27 . The method of  claim 21  wherein the preparation of said other antigenically similar species comprises spores, vegetative cells or combinations thereof. 
     
     
         28 . The method of  claim 21  wherein the species-specific monoclonal antibody is selected from the group consisting of IgA, IgE, IgG, IgM and associated sub-types. 
     
     
         29 . The method of  claim 21  wherein the host is selected from the group consisting of mice, rats, horses, cattle, chickens, sheep, goats, pigs and primates. 
     
     
         30 . The method of  claim 21  wherein boosting with the antigenically similar species is performed about seven days prior to fusing. 
     
     
         31 . The method of  claim 21  wherein boosting with  B. anthracis  is performed about three days prior to fusing. 
     
     
         32 . A species-specific monoclonal antibody to spores of  B. anthracis  made by the method of  claim 21 . 
     
     
         33 . A diagnostic kit comprising the antibody of  claim 32 . 
     
     
         34 . A hybridoma that expresses the antibody of  claim 32 . 
     
     
         35 . An antibody which is specifically reactive against  B. thuringiensis  and non-reactive against  B. cereus  or  B. anthracis.    
     
     
         36 . An antibody which is specifically reactive against  B. cereus  and non-reactive against  B. anthracis  or  B. thuringiensis.    
     
     
         37 . An isolated or recombinant antigen, or antigenically active portions thereof, comprising an EA1 protein of the surface layer of  B. anthracis.    
     
     
         38 . A pharmaceutical composition comprising the antigen, or active portions hereof, of  claim 37  and a pharmaceutically acceptable carrier. 
     
     
         39 . A method of using the antigen, or active portions thereof, of  claim 37  as the target for an immunological detection system for  B. anthracis.    
     
     
         40 . A vaccine against  B. anthracis  comprising a therapeutically effective amount of the antigen, or active portions thereof, of  claim 37 . 
     
     
         41 . A method for vaccinating against  B. anthracis  comprising administering the therapeutically effective amount of the vaccine of  claim 40  to a patient. 
     
     
         42 . A therapeutic agent comprising antibodies to the EA1 protein. 
     
     
         43 . A method for treating, preventing or controlling  B. anthracis  infection comprising administering an effective amount of the therapeutic agent of  claim 42  to a patient.