Monoclonal antibodies

Diagnostics Discovery Laboratory

Production of monoclonal antibodies

Production of monoclonal antibodies (mAb) is central to all efforts in the Diagnostics Discovery Laboratory (DDL). MAbs are monospecific antibodies that are exactly the same because they are made by identical immune cells that are clones of a unique parent cell. Georges Köhler, César Milstein and Niels Kaj Jerne shared the Nobel Prize in Physiology or Medicine in 1984 for the discovery of hybridoma technology.

Hybridoma Technology. MAb production begins with immunization of a mouse with an antigen. Several weeks after immunization, spleen cells are harvested. B cells in the spleen produce antibody but cannot grow in culture. The spleen cells are fused with cells of a myeloma line. The myeloma cells cannot produce antibody, but they will grow in culture. The two cells (spleen cells and myeloma cells) are fused together to produce a hybrid cell (hybridoma) that has the best properties of both cells—the hybridoma grows in culture and makes antibody.

Hybridoma cells are cloned to ensure that all cells in the culture are making exactly the same mAbs. Once cloned, the cells can be mass produced in tissue culture to make large amounts of the mAbs. Cell lines made in this manner are immortal; they can be frozen for years.

DDL has extensive experience in making mAbs to polysaccharides and proteins. These mAbs are the key reagents for diagnostics development. A core strength of DDL is use of proprietary strategies for immunization of mice and for screening of hybridomas to produce mAbs that are ideally suited for diagnostics development. mAbs that are well suited for other purposes such as therapeutics may not be well suited for the rigors of a lateral flow immunoassay. In another example, mAbs that function well in ELISA may not function well in LFA and vice versa.

Large scale production of mAbs. Diagnostics development requires large amounts of antibodies. The DDL uses two approaches to large scale antibody production. The CELLine two-compartment bioreactor is used for production of tens to hundreds of milligrams of mAb. This is usually sufficient for most purposes. The WAVE Bioreactor is used to produce gram amounts of mAbs. The WAVE Bioreactor is also used for production of recombinant proteins in CHO cells. The DDL is equipped with a WAVE Bioreactor™ 2/10 that allows for cell culture in amounts of 0.5-5 liters. Accessory equipment includes the PERFCONT2E optional perfusion module that allows for automated feeding and harvesting of cell cultures.