All tags Antibody guide A comparison between polyclonal and monoclonal

A comparison between polyclonal and monoclonal

Key differences, advantages, and disadvantages of polyclonal and monoclonal antibodies.

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Summary

Polyclonal antibodiesMonoclonal antibodies
Inexpensive to produceMore expensive to produce
Skills required for production are lowTraining is required for the technology used
Relatively quick to produceHybridomas take a relatively long time to produce
Generate large amounts of non-specific antibodiesGenerate large amounts of specific antibodies
Recognize multiple epitopes on any one antigen Recognize only a particular or restricted epitope on an antigen
Can have batch-to-batch variabilityOnce a hybridoma is made, it is a constant and renewable source

Low batch-to-batch variability


Polyclonal antibodies

Facts:

  • Recognize multiple epitopes on any one antigen. Serum obtained will contain a heterogeneous complex mixture of antibodies of different affinity.
  • Polyclonals are made up mainly of IgG subclass.
  • Peptide immunogens are often used to generate polyclonal antibodies that target unique epitopes, especially for protein families of high homology.

Antibody production:

  • Inexpensive and relatively quick to produce.
  • Production is less complex compared with monoclonal antibodies.


Advantages:

Polyclonal antibodies recognize multiple epitopes on any one antigen. This has the following advantages:

  • Polyclonals amplify the signal from a target protein with low expression level, as the target protein will bind more than one antibody molecule on the multiple epitopes. However, this is less advantageous for quantification experiments (eg in flow cytometry) as it generates inaccurate results.
  • More tolerant of minor antigen changes (eg polymorphism, heterogeneity of glycosylation or slight denaturation) than monoclonals.
  • Can identify proteins of high homology to the immunogen protein, and can be used to screen for the target protein in species other than that of the immunogen.
  • Often the preferred choice for detecting denatured proteins.


Disadvantages:

  • Prone to batch-to-batch variability.
  • Less useful for probing specific domains of an antigen because antiserum will usually recognize many domains.


Monoclonal antibodies


Facts:

Antibody production:

  • Training is required for the technology used.
  • Long timeframe for hybridoma production.


Advantages:

Hybridomas are a constant and renewable source once created, and all batches will be identical, increasing consistency and standardization of experimental procedures and results.

Monoclonals detect one epitope per antigen. This has the following advantages:

  • Monoclonals specifically detect a particular or defined/restricted epitope on the antigen, which means they are less likely to cross-react with other proteins.
  • Homogeneity of monoclonal antibodies is high. If experimental conditions are kept constant, results from monoclonal antibodies can be highly reproducible between experiments.


Disadvantages:

  • The epitope may not be shared across a range of species.
  • More vulnerable to the loss of epitope through chemical treatment of the antigen than polyclonal antibodies. This can be offset by pooling two or more monoclonal antibodies to the same antigen (eg cocktail antibodies).

See examples of cocktail antibodies from Abcam.

RabMAb® technology 

Abcam's RabMAb technology combines the benefits of both monoclonal and polyclonal technology to create highly specific and sensitive rabbit monoclonal antibodies.

Learn more about the advantages of RabMAb® technology.

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