Production of monoclonal antibodies.

Monoclonal antibodies are particularly specific for a single epitope and can be produced in large quantities. When immune splenic cells are fused with a tumor cell line, they produce hybridomas each secreting one highly specific antibody. These cells are then characterized and cloned to allow production over time. This unit describes the production of monoclonal antibodies beginning with basic protocols for immunization and cell fusion and selection. Support protocols are provided for screening primary hybridoma supernatants for antibodies of desired specificity, establishment of stable hybridoma lines, cloning of these B cell lines by limiting dilution to obtain monoclonal lines, and preparation of cloning/expansion medium (thymocyte-conditioned medium).