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The development of hybridomas has made antibodies invaluable for multiple applications including therapy, research, and diagnostics. Over the years, this innovative technology has been subsequently improved to increase the efficiency of the cell fusion process and stability of the resulting cell lines. Today, hybridomas are considered one of the most relevant technologies for antibody discovery and production. Check our frequently asked questions (FAQs) page about hybridomas for a complete overview of all steps of this robust process for antibody generation.
Hybridomas are immortal antibody-secreting cell lines first developed by scientists Georges Kohler and Cesar Milstein in 1975. Conventionally, these cell lines are generated by fusing short-lived spleen cells (plasma cells) from pre-immunized hosts with a compatible myeloma partner (malignant plasma cells).
This delicate process of cell fusion, often induced by electric pulses (electrofusion) or polyethylene glycol (PEG), renders these highly productive cell lines amenable to cryopreservation for the long-lasting production of high-quality monoclonal antibodies.
Besides this advantage, hybridomas allow:
These natural advantages of hybridomas make the technology invaluable for the discovery and production of antibodies for therapy, research, and diagnostic applications.
Hybridomas have played and continue to play a key role in antibody discovery for multiple applications. They allow capturing highly mature antibodies with the highest stability, affinity, and specificity towards specific targets.
These hybrid cell lines embody the possibility of indefinite production of monoclonal antibodies in vitro. Many of the highly valuable hybridoma cell lines are currently stored in vast cell banks allowing their widespread distribution and ultimately supporting research and diagnostic efforts.
The major sources of hybridomas are rodent species such as mice, rats, and hamsters. In these cases, monohybridoma production (fusion of a healthy plasma cell with malignant myelomas from the same species) is the most common route for achieving cell immortalization. However, some heterohybridomas (cell fusion between partners from different species) such as mouse-rat and rat-mouse are known to be as efficient as their monohybridoma counterparts.
Heterohybridomas have often been used to generate immortal antibody-secreting cell lines for alternative species such as chicken, rabbit, or human. However, most of these heterologous cell lines have proven to be genetically unstable and the process of clonal selection often leads to the loss of antibody-encoding genes.
After proper screening and monoclone selection, mouse hybridomas are able to secrete large quantities of IgG monoclonal antibodies. They can then be conjugated either to enzymes or fluorescent labels for the detection or capture of specific antigens.
This versatility is especially useful for diagnostics and research, where an antibody’s specificity and sensitivity may help to unravel the hidden causes of certain conditions or aid in the early-stage detection of many diseases. But in the long-term, hybridomas may suffer from instability resulting from mutations or chromosome loss that impact their long-term productivity.
To overcome this limitation, hybridoma cell lines can be sequenced so that the antibody-encoding genes may be easily adapted to recombinant expression in more stable mammalian systems.
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