Monoclonal antibodies production in vitro

Why produce in vitro?

The use of adjuvants such as Freund’s complete adjuvant or pristane has been a controversial issue since several years. The inflammation context they induce upon injection are potential sources of animal pain and distress. Therefore many countries have restricted their use and by extension the use of animals to produce monoclonal antibodies in vivo. Nevertheless, monoclonal antibody production can be performed in vitro by cell culture.

Thanks to adapted cell culture materials and well-established procedures, we can perform the production of monoclonal antibodies in vitro from milligram to gram scale without the constraint of animal handling.

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Hybridoma cells

You can require us to use a hybridoma clone we just developed for you within a monoclonal antibody development project. You can also provide us with your own clone in cell culture medium (shipped at 4°C), or frozen in cryotubes (shipped in dry ice). In the case of frozen cells, several additional weeks are required to expand the cells in the culture device. To be accepted for culture in our laboratory, please note that each hybridoma must be accompanied by a material transfert sheet as well as a certificate testifying the absence of mycoplasma.

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Procedure

Hybridoma cells are first expanded in culture flask or high density culture devices (for larger amounts of antibody) to reach the optimal cell density needed for the production.

Supernatant containing the monoclonal antibodies is then periodically collected until the expected quantity of antibodies is reached and can be subsequently purified on protein A or G column on request.

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Serum-free medium

Some immunological applications require very high quality antibodies and thus are not compatible with classic in vitro production methods which include fetal calf serum (FCS) in the medium. Our expertise in antibody engineering allows us to provide a service of in vitro monoclonal antibodies production without using FCS.

The use of serum-free medium shows several advantages as compared to classic media:

  • reproducible production: strict control over components composition, well defined medium, no variations due to changes of fetal calf serum batches,
  • improved specificity: no disturbance by natural calf IgG within the serum, better binding to protein A,
  • equivalent productivity: similar productivity in comparison with calf serum, no need for hybridomas to adapt to the environment,
  • accessible targets: antibodies directed against elements of the serum, no risk of aggregation.

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Yield

Average yield is about 20 μg/mL of monoclonal antibody in cell culture flasks, and can reach up to 1 mg/mL in a high cell density device.

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