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).
The therapeutic efficacy of antibodies is closely related to their ability to recognize and bind specific epitopes on target antigens. Epitopes, or antigenic determinants, are a group of amino acids or other chemical groups that are part of a molecule to which an antibody attaches itself. Epitope characterization can help reveal the mechanism of antibody binding and apply intellectual property (patent) protection for novel antibodies, in addition to designing antibodies with high specificity and minimal cross-reactivity.
Understanding the differences between antibody specificity and selectivity is essential for designing and interpreting antibody-based assays in research for experimental accuracy and data interpretation. Antibody specificity refers to an antibody's ability to recognize and bind to a particular epitope—a unique part of an antigen that elicits an immune response.
Antibody-based assays are essential tools in biomedical research, providing the means to detect, quantify, and visualize specific proteins or antigens within complex biological samples. These assays' efficacy hinges on the antibodies' precise properties. While affinity, avidity, specificity, and selectivity are fundamental to antibody performance, the ultimate impact of these properties is heavily influenced by the experimental context in which the antibody is employed.
Biologics, particularly antibodies, have become indispensable in biomedical research and therapeutic development. Research-use-only (RUO) biologics play a pivotal role in preclinical studies, providing researchers with the necessary tools to explore antibody functions and therapeutic potential in vivo.