Hybridoma Screening and Validation

Screening

Following fusion and HAT selection, hybridomas are screened by a high-throughput bead-based fluorescence analysis for the presence of IgG antibodies directed against the antigen. We also perform ELISA assays depending on the specific requirements of the screening strategy.

Cells from positive wells are expanded and cryostored to allow sufficient time for validation of the primary supernatant. We determine the IgG subclass and titer. These primary supernatants are delivered to our collaboration partners for further validation in the desired applications, e.g. WB, IP, IF, FACS, IHC, etc.

Usually, more than one clone is growing in a single well and primary supernatants originate from oligoclonal hybridomas rather than from a single monoclonal hybridoma. Therefore, we provide anti-IgG subclass-specific secondary antibodies (conjugated or unconjugated) that allow detection of the specific subclass and avoid false-positive results or high background.