Hybridoma Sequencing Service

Hybridoma Sequencing Service

Hybridoma cells are immortalized, but they are not stable. Mutations may occur during extensive passage, losing the entire antibody gene or specificity. Therefore, in the aspects of antibody manufacturing, patent application and downstream antibody engineering, antibody sequence information is very important for antibody quality. Tribiosciences has developed a PCR-based antibody sequencing workflow, combined with a degraded primer design. Compared with other methods such as mass spectrometry, this method is cost-effective and time consuming, and can sequence the full length and variable regions of the light and heavy chains of immunoglobulin (Ig) in hybridomas or B cells of mice, rabbits, alpacas, rats, humans, hamsters, humanized mice and other animals.

Steps of Antibody Sequencing

1. mRNA extraction from hybridoma cells or B cells
2. Reverse transcription of mRNA into cDNA
3. Amplification of antibody light and heavy chains
4. Clone light and heavy chain genes into a standard vector
5. DNA sequencing and annotation
6. Report antibody light and heavy chain sequences to the client