For the last few decades, antibodies have been thoroughly utilized as an extraordinarily powerful and important tool for the detection, quantification, characterization, and purification of molecules of interest. The foundation of their utility lies in a high degree of affinity and specificity to selectively bind a particular epitope present on carbohydrate, protein, or nucleic acid.
There are two primary types of antibodies used for research purposes, namely the polyclonal and monoclonal antibodies. Both the polyclonal and monoclonal antibodies have an important role in the immune system and can be utilized to answer different research needs. However, differences in their production techniques have given rise to several opposite characteristics between monoclonal and polyclonal antibodies.
This article aims to give an overview of their general production processes, differentiating properties, and their roles in research, diagnostics, and therapeutics purposes.
Polyclonal antibodies are a heterogeneous mixture of immunoglobulins (IgGs) that can recognize and bind to many different epitopes on the same antigen.
Polyclonal antibodies are produced by injecting a low amount of a particular antigen (proteins, synthetic peptides, whole organisms, cells, etc.) into an animal to trigger the development of different types of B cells and memory B cells to produce antibodies. After several days, the animal is given subsequent secondary or even tertiary booster immunization of the same antigen to produce higher titers of antibodies against the antigen.
Polyclonal antibodies can be obtained straight from the serum or purified to obtain a solution that is free from other serum proteins to limit the degree of cross-reactivity.
In contrast to polyclonal antibodies that are produced in vivo from several different B-cells in the animal, monoclonal antibodies are generated ex vivo using tissue culture techniques. Stable hybrid B-cell clones were cultured from a single parent cell — thereby generating an identical population of only a single type of IgG with monovalent affinity to target a particular epitope per antigen.
The production process begins immunization of the animal with the desired antigen multiple times. After several weeks, the B-cells are harvested from the animal’s spleen cells and immortalized by fusion with hybridoma cells (as spleen cells are unable to grow in culture) and then screened for the antibodies.
The hybridomas can grow continuously in culture while producing antibodies, thereby providing an unlimited source of identical monoclonal antibodies for mass production.
Polyclonal vs. Monoclonal: Advantages & Disadvantages
It is crucial to understand the differences and limitations of each process to choose the best fit for each specific application. The key advantages and disadvantages are listed in the table below.
Table 1: Advantages and disadvantages of polyclonal and monoclonal antibodies
|Polyclonal Antibodies||Monoclonal Antibodies|
|Advantages||● Relatively short production time (±3 months) without using complicated technologies
● More affordable
● High affinity and allows for quick binding due to recognition of multiple epitopes
● High sensitivity, even for proteins with low expression levels
● More tolerant to minor changes of antigen (slight denaturation, polymorphism, heterogeneity of glycosylation, etc.)
|● New animals will have to be immunized with the antigen to create a new batch (prone to batch-to-batch variability)
● Higher risk of background noise and cross-reactivity
|Disadvantages||● Highly specific recognition
● Possibility for large-scale production of identical antibodies
● High batch-to-batch reproducibility
● Minimal background noise and cross-reactivity
|● Long production time (±6 months) and requires high technical skills
● More expensive
● The necessity to produce a pool of several monoclonal antibodies
● Possible lower signal
● More vulnerable to minor changes of antigen (slight denaturation, polymorphism, heterogeneity of glycosylation, etc.)
Choosing the Right Antibody
The true potential of monoclonal antibodies lies in their monospecificity, homogeneity, and consistency. These attributes highly suggest their use in clinical settings, affinity purification, and quantitative detection experiments. However, the technology used for the production of monoclonal antibodies of the desired specificity is frequently difficult, expensive, and extremely time-consuming.
Several suggested applications of monoclonal antibodies include:
- Therapeutic drug development
- Flow cytometry
- X-ray crystallography
- Gene sequencing.
For general research applications, the advantages of polyclonal antibodies typically outweigh the advantages that monoclonal antibodies provide. Polyclonal antibodies can be generated much more rapidly, at a much lower price, and with less complicated technical skills than is required to produce monoclonal antibodies.
Polyclonal antibodies are suitable for a wide range of applications, including:
- Hemagglutination reactions
- Immunoprecipitation assays
- ELISA, WB, microarray assays, IHC, etc.
- IP or ChIP.
Custom Rabbit Polyclonal Antibody Production Service
There is a large choice of host species to choose from for your antibody generation. However, rabbit is the common host choice for producing polyclonal antibodies. Rabbit is used 95% of the time as it has a bigger repertoire for antigen classes, and generally provides the greatest yields in the shortest period of time (1-5 mg when purifying a 20 mL serum).
Antibodies production is a technically demanding process that requires in-depth knowledge and monitoring capability achievable by an average laboratory. Although there are readily available reagents that propose quick solutions and eliminate the issues of time and cost, these products do not always fulfill the requirements of particular research studies.
In this case, opting for custom rabbit polyclonal antibody production and development services can offer added security, assurance, and control of your scientific success.
With rich experience as one of the world’s leading antibody manufacturers, Boster Bio offers an affordable, robust, and flexible custom rabbit polyclonal antibody production service to help you manage your polyclonal antibody development project from antigenic selection to delivering affinity purified antibodies. Boster’s custom rabbit polyclonal antibody service platform can develop high-quality custom rabbit polyclonal antibodies for your downstream applications!