Methods of producing hybridomas and monoclonal antibodies and antibodies produced thereby

Abstract

Disclosed are a method of producing a hybridoma, a method of making an antibody involving the hybridoma, and an antibody made by the method.

1 . A method of producing a hybridoma, comprising:
 (a) inoculating a mouse by subcutaneous, intradermal, or intramuscular injection with a recombinant adenoviral vector that comprises a gene encoding a protein of interest;   (b) isolating B-cells from the mouse about 10 days post-inoculation;   (c) fusing the isolated B-cells with mouse myeloma cells to produce a plurality of hybridomas;   (d) identifying a hybridoma from (c) that produces a monoclonal antibody that specifically binds the protein of interest.   
     
     
         2 . The method of  claim 1 , wherein the mouse is a C57B1/6 mouse or non-obese diabetic (NOD) mouse. 
     
     
         3 . A method of making an antibody, comprising the method of  claim 1 , and further comprising isolating the antibody from the hybridoma of (d). 
     
     
         4 . The method of  claim 3 , further comprising producing the antibody by recombinant expression of its amino acid sequence. 
     
     
         5 . An antibody made by the method of  claim 3  or  claim 4 .