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Polyclonal Versus Monoclonal Antibodies
Polyclonal and monoclonal antibodies are among the most widely used reagents for scientific research and are named according to how they are produced. While a polyclonal antibody preparation comprises multiple antibodies that each recognize a different epitope, a vial of monoclonal antibodies contains just one type of antibody that is derived from a single B cell clone. Both polyclonal and monoclonal antibodies have distinct features and benefits which should be considered during antibody selection.
How are polyclonal and monoclonal antibodies produced?
Irrespective of whether an antibody is polyclonal or monoclonal, its production begins by immunizing an animal with the protein target of interest. This is taken up by antigen presenting cells (APCs) and displayed at the cell surface, allowing its recognition by circulating B cells. When a B cell encounters its target antigen and binds via the unique antibody expressed at the B cell surface, it undergoes clonal expansion to yield a pool of identical B cells, each producing antibodies that can be purified from the sera. Because any protein used for immunization will have multiple epitopes, the purified antibody preparation will contain antibodies from multiple B cell clones, hence the term polyclonal.
Monoclonal antibody production instead involves isolating the B cells from the immunized animal and fusing them with immortal myeloma cells to generate hybridoma. By culturing these such that each well of a multi-well plate receives just one hybridoma - which can then be expanded and tested - it is possible to select the most specific clones for future use. Since every clone will produce just a single type of antibody that can be extracted from the cell culture supernatant for research use, the resultant product is referred to as being monoclonal. More recently, recombinant methods for generating monoclonal antibodies have been developed that circumvent the need for animal use.
What are the advantages and disadvantages of polyclonal antibodies?
A main advantage of polyclonal antibodies is their capacity for signal amplification, which results from multi-epitope recognition and can increase the likelihood of detecting a scarce target. Polyclonal antibodies may also be available sooner when a new target (e.g., SARS-CoV-2) is discovered since they are relatively quick and easy to produce. Another important benefit of polyclonal antibodies relates to the animal species used for production. Polyclonals are typically produced in goats, which allow antibodies to be generated in large volumes, or rabbits, which have a more diverse immune repertoire than mice (the major species used for hybridoma production) and can yield antibodies against challenging targets, such as small peptide antigens.
Disadvantages of polyclonal antibodies are that they are available in only limited supply, corresponding to the lifespan of the host animal, and can demonstrate variable performance from lot-to-lot if not carefully validated. Polyclonal antibodies may also present an increased risk of non-specific binding, although many antibody manufacturers address this by performing cross-adsorption.
Our rigorously validated HRP and Alexa Fluor conjugated secondary antibodies are cross-adsorbed against multiple species and include products for detecting mouse, rabbit, goat, human, and chicken IgG.
What are the advantages and disadvantages of monoclonal antibodies?
Monoclonal antibodies offer the advantage of high specificity, which is achieved by screening and selecting individual clones during antibody development. They also demonstrate consistent performance and are available in unlimited supply, with their repeat production avoiding any need for further animal use. Although monoclonal antibodies have historically been more expensive than polyclonal antibodies, this is less frequently the case.
| Polyclonal | Monoclonal | |
|---|---|---|
| Advantages | May provide signal amplification due to epitope redundancy
Often available sooner than monoclonal antibodies
Can be produced for challenging targets such as small peptide antigens
| High specificity
Consistent performance, ensuring more reproducible results
Unlimited supply
Reduced animal use compared to polyclonal antibody production
|
| Disadvantages | Limited supply, corresponding to the lifespan of the host animal
Lot-to-lot variability if not carefully validated
Potential for non-specific binding
| Can take longer to reach the market
May be more expensive
|
How do I decide whether to use a polyclonal or a monoclonal antibody?
Both polyclonal and monoclonal antibodies are used extensively for techniques such as Western blot, immunohistochemistry (IHC), flow cytometry, ELISA, and immunocytochemistry (ICC). Deciding which antibody product best suits your needs depends largely on the aim of your research and you may choose to combine polyclonal and monoclonal antibodies in the same experiment. Provided you select well-characterized antibodies that have been proven to detect the target of interest in your chosen species and application, you'll be off to a strong start.
We offer a wide selection of polyclonal and monoclonal antibodies, including products specific to human, mouse, and rat isotypes and subtypes.
