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).
Antibodies are versatile molecules that perform a range of effector functions, many of which engage different arms of the immune system. Their modes of action extend beyond simple antigen binding, enabling the activation of various immune mechanisms that lead to pathogen neutralization and clearance. These functions include blocking molecular interactions, activating the complement system, and linking the humoral immune response to cellular immune responses via Fc receptor engagement.
In today’s competitive biotech landscape, intellectual property (IP) protection has become an essential pillar in fostering innovation and collaboration across drug discovery and development. By offering clear IP terms and no royalty fees,pharmaceutical companies and research institutes
In addition to isotypes and subtypes, antibodies exhibit genetic variation known as allotypes, which are polymorphic epitopes on immunoglobulins. These allotypic differences arise from allelic variations in immunoglobulin genes, causing certain antibody subtypes to differ between individuals or ethnic groups. The presence of these polymorphic forms can influence immune responses, particularly when an individual is exposed to a non-self allotype, potentially triggering an anti-allotype immune reaction.
In mammals, antibodies are classified into five major isotypes: IgA, IgD, IgE, IgG, and IgM. Each isotype is defined by the heavy chain it contains: alpha (IgA), delta (IgD), epsilon (IgE), gamma (IgG), or mu (IgM). These structural differences in the heavy chains determine the antibody's function, tissue localization, and role in the immune response. Furthermore, antibody light chains fall into two classes—kappa and lambda—with kappa being more common, though both exhibit similar functions despite differences in sequence.
