polyclonal vs. monoclonal

About polyclonal antibodies

Advantages

Polyclonal antibody development requires moderate technical skills when compared to a monoclonal antibody. As soon as the immunisation protocol has been carried out successfully, polyclonal antibodies contained in the serum are indeed ready-to-use. Further purification can also help improve affinity and specificity.

The multi-epitope specificity of polyclonal antibodies makes them useful in detection and capture applications, such as IP and ChIP for which these antibodies give better results. Besides, lower amounts of proteins can be detected, for one molecule of antigen may bind more than one antibody.

Upon denaturation, although some epitopes may be lost due to conformational changes, the probability that one or more epitopes remain intact and bind to antibodies is still high. Development protocols for polyclonal antibodies are well-proven and relatively short (down to 2 months) when compared to monoclonal antibodies.

Disadvantages

Multiple epitope recognition often induces high cross-reactivity with proteins containing conserved patterns, especially with inter-species homologous proteins. Using a peptide as immunogen can help circumvent this problem and increase the specificity of your polyclonal antibody. The presence of high quantity of non-specific antibodies often leads to high background levels. Purifying polyclonal antibodies by antigen affinity chromatography also helps increase the specificity of your antibody and thus decrease this false positive signal.

Production of polyclonal antibodies is animal-dependent and may vary dramatically from one individual to another.

Frequently asked questions about polyclonal antibodies

Back to top

About monoclonal antibodies

Advantages

If cell culture conditions are kept stable, the production of monoclonal antibodies can be almost indefinitely renewable. With adapted cell culture equipment, large-scale production can provide up to milligrams of antibody within a week. Stable production provides a unique homogeneity feature to these antibodies. Such uniform tools lead to more reproducible experiments, and are then often used as standards.

As their specificity is very high for their epitope, monoclonal antibodies show very low background staining when compared to polyclonal ones. Additionally, unless their epitope is a conserved pattern, monoclonal antibodies have reduced risks of unexpected cross-reactivity.

Disadvantages

Nevertheless, developing a monoclonal takes time and requires high technical skills. Development protocols are more than three times as long as for polyclonal antibodies, for the generation of stable hybridomas and isolation of clones requires several cycles of cell expansion in vitro.

Moreover, as the epitope is unique among the population of monoclonal antibodies, even a slight change in conformation may lead to dramatically reduced binding capacity. Careful considerations should be taken especially if your experiments involve denaturing conditions.

Frequently asked questions about monoclonal antibodies

Back to top

◁ Back to Antibody Customisation page

Newsletter Receive news, promotions and offers in your e-mail box