Method for making monoclonal antibodies and cross-reactive antibodies obtainable by the method

Abstract

A method of making monoclonal antibodies according to a mixed antigen immunization protocol is described. In addition, antibodies obtainable by the method are disclosed which specifically cross-react with two or more different receptors to which Apo-2 ligand (Apo-2L) can bind.

1 . An isolated nucleic acid comprising DNA encoding an antibody which specifically cross-reacts with two or more different Apo-2L receptors. 
     
     
         2 . The nucleic acid of  claim 1  wherein the antibody comprises a monoclonal antibody. 
     
     
         3 . The nucleic acid of  claim 1  wherein the antibody specifically binds to Apo-2 polypeptide and further specifically cross-reacts with another Apo-2L receptor. 
     
     
         4 . The nucleic acid of  claim 1  wherein the antibody specifically binds to Apo-2 polypeptide and further specifically cross-reacts with DR4. 
     
     
         5 . The nucleic acid of  claim 1  wherein the antibody is an agonistic antibody. 
     
     
         6 . The nucleic acid of  claim 1  wherein the antibody is a blocking antibody. 
     
     
         7 . The nucleic acid of  claim 1  wherein the antibody is an antibody fragment. 
     
     
         8 . The nucleic acid of  claim 1  wherein the antibody comprises non-human hypervariable region residues and human framework region residues. 
     
     
         9 . The nucleic acid of  claim 1  wherein the antibody is a human antibody. 
     
     
         10 . The nucleic acid of  claim 1  wherein the Apo-2L receptors are native sequence Apo-2L receptors. 
     
     
         11 . The nucleic acid of  claim 5  wherein the agonistic antibody binds to Apo-2 polypeptide or DR4. 
     
     
         12 . An isolated nucleic acid comprising DNA encoding an antibody having the biological characteristics of a monoclonal antibody selected from the group consisting of 3H1.18.10 (produced by the hybridoma having ATCC Accession No. HB-12535), 3H3.14.5 (produced by the hybridoma having ATCC Accession No. HB-12534) and 3D5.1.10 (produced by the hybridoma having ATCC Accession No. HB-12536). 
     
     
         13 . The nucleic acid of  claim 12  wherein the antibody binds to the same epitope as the epitope to which a monoclonal antibody selected from the group consisting of 3H1.18.10 (produced by the hybridoma having ATCC Accession No. HB-12535), 3H3.14.5 (produced by the hybridoma having ATCC Accession No. HB-12534) and 3D5.1.10 (produced by the hybridoma having ATCC Accession No. HB-12536) binds. 
     
     
         14 . The nucleic acid of  claim 12  wherein the antibody has the hypervariable region residues of a monoclonal antibody selected from the group consisting of 3H1.18.10 (produced by the hybridoma having ATCC Accession No. HB-12535), 3H3.14.5 (produced by the hybridoma having ATCC Accession No. HB-12534) and 3D5.1.10 (produced by the hybridoma having ATCC Accession No. HB-12536). 
     
     
         15 . A vector comprising the nucleic acid of  claim 1 . 
     
     
         16 . A host cell comprising the nucleic acid of  claim 1 . 
     
     
         17 . A method of producing an antibody comprising culturing the host cell of  claim 16  under conditions wherein the DNA is expressed. 
     
     
         18 . The method of  claim 17  further comprising recovering the antibody from the host cell culture. 
     
     
         19 . The method of  claim 18  further comprising combining the recovered antibody with a pharmaceutically acceptable carrier. 
     
     
         20 . The method of  claim 18  further comprising conjugating the recovered antibody with a heterologous molecule. 
     
     
         21 . The method of  claim 20  wherein the heterologous molecule is polyethylene glycol, a label or a cytotoxic agent.