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Production of Monoclonal Antibodies

Hybridoma Technology: Production of Monoclonal Antibodies 


Monoclonal antibodies (mAbs) are monovalent (one epitope) antibodies derived from clones of a single parental immune cell. The original method for producing monoclonal antibodies is found in hybridoma technology. Prior to the production of monoclonal antibodies, polyclonal antibodies (multiple epitopes, multiple parental cells) were the standard for in vitro research. Production of polyclonal antibodies, however, was too inconsistent to be an effective tool for in vivo research. The breakthrough came in the 1970s when Kohler and Milstein succeeded in creating the first hybridoma cell line. They were awarded the Nobel Prize for Medicine and Physiology in 1984.


Hybridoma technology is a common method used for the production of monocolonal antibodies. The method of in which antibody-producing B cells are fused with immortal cancerous cell lines was developed by Kohler and Milstein. It involves immunizing a host animal with an antigen of interest to elicit an immune response. When a sufficient level of response is achieved as determined by in vitro testing, the mature B-cells are harvested from the animal spleens. The B-cells are then fused with myeloma cells using polyethylene glycol (PEG) to generate an immortalized hybridoma cell line. Through multiple rounds of screening, a single, pure, hybridoma positively expressing the antibody against the antigen of interest, as determined by ELISA, is amplified and monoclonal antibodies are purified.


Hybridoma technology has enabled the development of many types of monocolonal antibodies. Due to their high target affinity, monoclonal antibodies have a wide range of diagnostic and therapeutic uses. They are employed in the detection of the target antigen/protein in ELISAs, Western Blots, and in tissue sections inimmunohistochemistry. Monoclonal antibody therapy is employed in the prevention and/or treatment of infection by Respiratory Syncytial Virus (RSV) and Ebola or as immunotherapeutics for such illnesses as rheumatoid arthritis, and cancers.

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Fig. 1 Survival after challenge with INFV H1N1 A/Pert/261/2009 (Tamiflu-resistant strain). Inoculum 1xLD90=1.0E+05 PFU/mouse
Survival after challenge with INFV H1N1 A/Pert/261/2009 (Tamiflu-resistant strain) 1.0E+05 PFU/mouse
Survival and weight change in BALB/c mice challenged with INFV A/ Texas/36/91 (H1N1) and treated with antiviral Osletamivir Phosphate (Tamiflu)
Lung viral load and Survival (30 % weight loss cut-off) in BALB/c mice challenged with INFV H3N2 A/HK/1/68.