Seeding of selected hybridomas

Seeding the selected hybridomas

At this stage, the selected hybridomas are still polyclonal. To isolate clones from each other, each selected hybridoma is seeded into 96-well plates using the limiting dilution method, and cultured for two weeks.

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Identification of growing clones

Screening of growing cell lines

A visual control is then performed to check for isolated cell clusters which correspond to one single clone. After another week of growth, the supernatant of cell cluster-containing wells are assayed by ELISA to identify the clones which specifically produce the expected antibody. Based on our results, we select up to six positive clones per hybridoma.

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Confirmation of antibody production


The cells from the corresponding wells are then transferred into a 24-well culture plate and expanded for one week. The supernatants are then assayed again to check for stable antibody production. We also ship to you 1 mL of each supernatant to run tests in your specific conditions.

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It's up to you!


Based on your results, you select one final clone for each selected hybridoma. The cells are then expanded in 25 cm2 flasks for freezing. A complete isotyping is performed to determine the class and subclass of the antibody as well as the isotype of the light chain.

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