Monoclonal Antibody Production

Monoclonal antibodies are specific against a single epitope on the antigen unlike polyclonal antibodies, which are a mixture of antibodies against various epitopes of an antigen. This specificity of monoclonal antibodies allows researchers to probe for that epitope even when the changes in that epitope are subtle—sometimes as small as a single amino acid change. A major advantage of the monoclonal antibodies is because they are generated from either a hybridoma or recombinant cell line, there will be an unlimited supply of these antibodies with the same specificity and structure.

Labcorp utilizes two different technologies to generate monoclonal antibodies: hybridoma and B-cell cloning. For hybridoma technology, rodents are immunized with the antigen and when a sufficient immune response is detected, antibody-producing cells are fused with a myeloma cell line and screened. Alternatively, in B-cell cloning, the splenocytes from immunized animals are screened and the heavy and light chain genes are cloned and expressed from selected B-cells. Both technologies provide the opportunity to generate a nearly unlimited supply of antibodies with identical properties.

Hybridoma generation

• Rodent host species
• Splenocytes and/or lymph nodes used for fusion
• Various fusion procedures such as polyethylene glycol (PEG) or electrofusion
   

B-cell cloning

•  Nearly any host species
• Antigen-specific B-cell isolation by FACS
• Cloning of heavy and light chain genes
• Expression in mammalian vectors and stable cell generation
   

Antibody purification and characterization

• Scale-up:
  •  Ascites or in vitro methods
  • Mycoplasma testing, isotyping
• Cell banking/storage of hybridoma cell lines
• Transient transfection of mammalian vectors