Monoclonal antibodies (monoclonal antibodies) are highly specific antibodies that are used in areas such as disease diagnosis,** and research. The following is the production and preparation process for monoclonal antibodies:
Antigen selection: Determine the target antigen and obtain the purified target antigen.
Immunized animal immunization: The target antigen is injected into the body of an immunized animal (usually a mouse) to trigger an immune response. Through multiple immunizations, immune cells can be stimulated to produce antibodies specific to various antigens.
Hybridoma technology is performed: immune cells (B cells) and tumor cells from immunized animals are extracted and fused to form hybridoma cells. Hybridoma cells have the longevity and antibody-producing capacity of immune cells, as well as the infinite replication capacity of tumor cells.
Cloning and selection: Hybridoma cells are monoclonalized, i.e., single cells are isolated and cultured to form monoclonal cell populations. Then, monoclonal cell lines that produce target antibodies are screened by methods such as enzyme-linked immunosorbent assay (ELISA).
Culture and collection: Monoclonal cell lines are cultured in vitro to produce a large number of monoclonal antibodies. Subsequently, the cell culture medium (supernatant) is collected.
Purification and concentration: Purification and concentration of monoclonal antibodies are carried out by protein ag affinity chromatography, gel filtration, ion exchange chromatography and other technologies. These methods can remove other impurity proteins and improve the purity of monoclonal antibodies.
Determine mAb activity: Verify the specificity and activity of purified mAbs using various methods (e.g., Western blot, ELISA).
Preparation and storage: Monoclonal antibodies are formulated, filter-sterilized, and then aliquoted and stored for subsequent applications.
It is important to note that the production and preparation of monoclonal antibodies is complex and requires suitable laboratory conditions. In addition, there are emerging technologies, such as recombinant antibody technology and monoclonal antibody humanization, to improve the efficacy and stability of monoclonal antibodies.