Monoclonal Antibody Production

The traditional monoclonal antibody (mAb) production process usually starts with generation of mAb-producing cells (i.e. hybridomas) by fusing myeloma cells with desired antibody-producing splenocytes (e.g. B cells). These B cells are typically sourced from animals, usually mice. After cell fusion, large numbers of clones are screened and selected on the basis of antigen specificity and immunoglobulin class. Once candidate hybridoma cell lines are identified, each "hit" is confirmed, validated, and characterized using a variety of downstream functional assays. Upon completion, the clones are scaled up where additional downstream bioprocesses occur.

A typical monoclonal antibody production process.

A typical monoclonal antibody production process

  1. Immunization of mice & isolation of splenocytes - Mice are immunized with an antigen and later their blood is screened for antibody production. The antibody-producing splenocytes are then isolated for in vitro hybridoma production.
  2. Preparation of myeloma cells - Myeloma cells are immortalized cells that, once fused with spleen cells, can result in hybridoma capable of unlimited growth. Myeloma cells are prepared for fusion.
  3. Fusion - Myeloma cells and isolated splenocytes are fused together to form hybridomas in the presence of polyehthylene glycol(PEG), which causes cell membranes to fuse.
  4. Clone screening and picking - clones are screened and selected on the basis of antigen specificity and immunoglobulin class.
  5. Functional characterization - Confirm, validate and characterize (e.g. ELISA) each potentially high-producing colony.
  6. Scale up and wean - Scale up clones producing desired antibodies and wean off selection agent(s).
  7. Expansion - Expand clones producing desired antibodies (e.g. bioreactors or large flasks).

For more information, please download our eBook, "Select high-value cell lines like a pro".

Other links: