Best Practices for Screening Hybridoma Clones
Hybridomas are inherently unstable; screening them to identify clones that exhibit optimal growth and antibody yield is an essential part of establishing stable clones. The ClonaCell™-HY product line includes media for hybridoma HAT selection, and expansion media to help streamline the hybridoma development process.
If you are screening hybridoma cultures, you can ensure that your resources are used for expanding only stable clones by following these best practices:
- Choose an established screening method. Some commonly used methods for screening hybridomas include flow cytometry, ELISA, and western blot.
- Perform a preliminary screening step and expand the positive hybridomas. This should be followed by a second round of confirmatory tests to ensure stable antibody production by the preliminary positive hybridomas. You may observe a different result in the confirmatory tests. This could be due to the unstable nature of the clones.
- Use species-specific secondary antibodies. It is a good practice to include a positive control in the screening process by using a purified antibody bound to a target antigen.
- Cryopreserve positive hybridomas as a backup source while expanding clones.
- Sub-clone positive hybridomas as soon as possible. If the culture is non-clonal, this will ensure that non-expressing cells do not overgrow the antibody-producing hybridoma cells.
How to Generate Hybridomas Using Semi-Solid Cloning
Watch a video outlining the steps in selection and growth of mouse hybridomas using ClonaCell™-HY.
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