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Hybridoma Sequencing

Biointron 2024-01-26 Read time: 2 mins
Image credit: DOI: 10.2289/fmicb.2017.00495

Hybridoma technology revolutionized the field of antibody drug discovery by enabling the mass production of monoclonal antibodies to an antigen of interest. Hybridoma cell lines are created by fusing antibody-producing B cells with immortalized myeloma cells, resulting in cells that produce specific monoclonal antibodies. However, these cell lines present certain limitations, including contamination risks, low yield, and storage space constraints. To address these challenges, hybridoma sequencing emerges as a powerful tool that not only overcomes these disadvantages but also prevents the loss of hybridoma cell lines.

Hybridoma sequencing is the sequencing of the variable heavy (VH) and variable light (VL) domains of monoclonal antibodies produced from your hybridoma cell line. Your candidate antibody can always be manufactured via recombinant expression after sequencing with mammalian cells, such as HEK/CHO. In addition, sequencing hybridoma cells allows us to produce humanized antibodies, as well as authentication of a hybridoma cell line. The process is as follows:

  1. Hybridoma cell line delivery and validation

  2. mRNA extraction

  3. Reverse transcription

  4. scFv sequencing by PCR amplification

  5. Sub-cloning of variable domains into a standard vector

  6. DNA sequencing of at least 5 clones for sequence alignment

  7. Sequencing data analysis

  8. Optional: antibody expression

Biointron's hybridoma sequencing service offers competitive pricing, fast turnaround times (1 week to deliver sequencing results after receiving the hybridoma cell line), and guaranteed 100% sequence accuracy (cross-verified with five independent clones).

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