Cloning of selected hybridomas

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.

Back to top

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.

Back to top

Confirmation of antibody production

ELISA

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.

Back to top

It's up to you!

Validation

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.

Back to top

◁ Back to Antibody Development page

Newsletter Receive news, promotions and offers in your e-mail box