Recently, rabbit monoclonal antibody formation via phage display has become revered amongst researchers and manufacturers worldwide as an alternative to antibodies from rodents. These antibodies can be used in multiple scenarios, including flow cytometry, ELISA, western bot, immunofluorescence, and immunohistochemistry.
However, before examining the process of rabbit monoclonal antibody formation and their advantages, we’ll take a closer look at antibodies and why they’ve become so important in scientific research.
What Is an Antibody?
Antibodies are simply proteins dispelled by B cells present in the immune system. These proteins are crucial in detecting, binding, and dissipating the effects of antigens tied to specific organisms, including viruses and bacteria.
The importance of antibodies can’t be overemphasized. They’re the bedrock of scientific research and diagnosis due to their ability to detect and identify antigens. The attribute of an antibody to seamlessly bind with an antigen is highly dependent on the latter’s surface — the epitope. Binding success also depends on how well the antigen’s surface can attach itself to the antibody.
Antibodies are shaped in the form of a Y.
The base is responsible for what’s tagged the effector function. The arms or branches are responsible for seamless antigen detection.
Monoclonal or polyclonal antibody creation lies with the ability of B cells in a host to stimulate and produce antibodies at a steady rate. Created antibodies are harvested and purified to facilitate scientific and clinical use.
Notable Differences Between Polyclonal Antibodies and Monoclonal Antibodies
When an animal is injected with an antigen, the B cells inherent in its system start to secrete various antibodies to combat and bind the epitopes present in the immunogen (antigen). The antibodies produced at this point are referred to as polyclonal antibodies.
Conversely, monoclonal antibodies are engineered to be reproduced from clone B cells from the host animal. Thus, these antibodies can only attach themselves to similar antigens, thus reducing the cross-activity risks.
While polyclonal antibodies are produced in what’s known as in vivo, monoclonal antibodies are developed in an ex vivo format. Although both antibody variations have advantages and disadvantages, their heterogeneity greatly limits polyclonal antibodies.
Conversely, monoclonal antibodies have become popular for their use in multiple applications.
Why Are Rabbit Monoclonal Antibodies Preferred?
Compared to common animal mediums (rats and mice), rabbits rank as a suitable media for monoclonal antibody development due to their immune system being responsive to various antigens. Physically, rabbits are also the preferred alternative as they have larger spleens — an organ that generates more antibodies.
The Process of Rabbit Monoclonal Antibody Production
Rabbit monoclonal antibodies share some semblance with those gotten from mice. However, the latter is preferred due to its higher sensitivity and specificity rates. The spleen cells obtained from immunized rabbits are combined with partner cells to create an unbeatable cell line that can form more antibodies.
The antibodies derived from a single clone are characterized to understand their performance in multiple applications. After this screening process, the best clone is sieved out for antibody production.
Advantages of Rabbit Monoclonal Antibodies
The rabbit’s immune system is unique and can yield the following advantages when creating monoclonal antibodies:
- High Affinity and Specificity
Rabbit monoclonal antibodies are efficient and effective for seamlessly recognizing minor epitope formats and combining target protein variations. Moving on, rabbit monoclonal antibodies have incredibly low equilibrium dissociation constants (KD). KD, in this context, occurs at a picomolar range where it equals 10 to 12M.
- Reduced Background Noise
Rabbit customs and antibodies can only decipher one epitope. Thus, the chances of them conflicting with other proteins are almost non-existent. Since rabbit monoclonal antibodies combine with antigens swiftly, they have an increased signal-to-noise ratio than antibodies collected from a mouse or rat.
- Distinctive Epitope Recognition Levels
Compared to a mouse’s immune system, the average rabbit’s immune system has lesser immunodominance and wider B-cell variety. These attributes guarantee increased epitope detection rates and chances of detecting novel epitopes while creating rabbit monoclonal antibodies.
- Suitable for IHC
Immunohistochemistry, otherwise known as IHC, is important to certify monoclonal antibodies’ tissue distribution concerning illness and health. For antibodies to work well for IHC, they must be able to detect epitopes in a noisy background. Rabbit monoclonal antibodies are useful for this endeavor due to their sensitive yet specific nature, even in the face of challenging applications like the IHC.
How About Recombinant Rabbit Monoclonal Antibodies?
Recombinant antibody protocol is now being used to develop rabbit monoclonal antibodies. This technique used in rabbit antibody creation yields higher sensitivity, specificity, and performance levels.
Monoclonal rabbit antibodies created through the recombinant antibody technique are similar to regular monoclonal rabbit antibodies in hybridomas. The only disparity between both antibody types is that the latter isn’t susceptible to lot-to-lot variation and cell line drifting.