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:
Hybridoma cell line delivery and validation
mRNA extraction
Reverse transcription
scFv sequencing by PCR amplification
Sub-cloning of variable domains into a standard vector
DNA sequencing of at least 5 clones for sequence alignment
Sequencing data analysis
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).
Antibody specificity refers to an antibody's ability to selectively bind to a unique epitope on a target antigen while avoiding interactions with unrelated antigens. This property arises from the highly specialized antigen-binding site located in the variable region of the antibody, which determines its unique binding characteristics.
Antibody affinity refers to the strength of the binding interaction between a single antigen epitope and the paratope (binding site) of an antibody. This interaction is a fundamental measure of how well an antibody recognizes its specific antigen target.
Recombinant antibodies are produced using genetic engineering techniques, unlike traditional antibody production, where the immune system generates antibodies without direct control over their sequence. By introducing genes encoding antibody fragments into host cells, such as bacteria or mammalian cells, recombinant antibodies can be expressed, purified, and deployed for applications including research, diagnostics, and therapeutics.
Recombinant antibody expression is a biotechnological process that involves engineering and producing antibodies outside their natural context using recombinant DNA technology.