Many people enter immunobiology from a different area of research. Although they swiftly learn the protocols and methods of the assays, they may not fully understand the underlying concepts. We at Novus Biologicals realise that not everybody purchasing products from our antibody catalogue has an in-depth knowledge of the “tools of the trade”. For them, and anybody else needing a quick bit of revision, we have put together a few basic notes.
Most people quickly understand that antibody proteins are produced by B lymphocytes, and are specific to foreign proteins (antigens) that enter the body. The body’s natural ability to produce such antibodies has been utilised into a useful research tool. Synthetic and natural antigens are used to generate antibodies, which are then used as probes to detect and bind to those target antigens in a variety of research applications.
The generating, modifying and purifying procedures that were used to create the first antigen probes established definitive protocols which remain little changed today. Hart and Lane’s Antibodies: A Laboratory Manual was published in 1988 but is still the industry bible. Now, as then, purified antigens injected into lab or farm animals evoke high-level expression of antigen-specific immunoglobulins, which are then harvested and purified.
This method produces heterogeneous polyclonal antibodies. Although they all respond to the same antigen, they are derived from many different B-cell lines, each of which recognises a specific epitope (amino acid sequence) on the protein. These are useful for tagging studies, but there is a need for monoclonal immunoglobulins as well. These are produced by immunising mice as above, removing the spleens, and then fusing the B-cells with immortal myeloma cell lines. This results in antibodies specific to a single epitope on the antigen.
