As a world leader in antibody production, ProteoGenix strives to offer the best of antibody production to its customers. Our wish is to link the most ambitious and innovative antibody production projects with our highly skilled and passionate team of experts. Our expertise and state-of-the-art technologies coupled with the enthusiasm we daily put in succeeding in your projects explains our unrivalled track record.
Our antibody production services
Choose a partner having more than 15 years of experience in high-quality polyclonal antibody production. ProteoGenix offers one-stop solution from antigen design to purified polyclonal antibody delivery.
You can even order online thanks to our entertaining interactive form!
Since 2003, ProteoGenix has been striving to accompany scientists in their monoclonal antibody production. Our scientists guide you from monoclonal antibody generation by hybridoma development or antibody phage display to bioproduction. Our monoclonal antibody production services are fully custom and can include a wide range of engineering possibilities such as antibody humanization, affinity maturation…
A doubt about the most adapted antibody production technique for your project? Polyclonal antibodies or monoclonal antibody? Hybridoma or Phage display? Get a clear and personalized answer in less than two minutes thanks to our innovative online tool.
Try our tool to choose the best technique
for your antibody production
As a global leading antibody production company, ProteoGenix strives not only at providing best-in-class services but also a broad range of solutions to carefully adapt to each project specifications. Our passionate team is also willing to share its long-standing experience with its customers. In consequence, we developed a new interactive form to guide you to define the most relevant antibody production strategy for your project.
Why choose ProteoGenix’s custom
antibody production services?
A service adapted to your application
Our extended and unique portfolio of antibody production services allows us to propose solutions to all your challenges.
ProteoGenix is committed in animal well-being in science by applying the highest ethical standards.
Unique guarantees market
Our work is guided by customer satisfaction and explains our unrivalled success rate. It allows us to offer the highest guarantees on the market.
PhD account managers
We put at your disposal highly skilled account managers who guide you along your decision-making process.
Various antigen design approaches
Trust our experienced team to propose the most adapted antigen design strategy to get the most effective antibodies.
Therapeutic, diagnostic, research… As an antibody production company, ProteoGenix offers one-stop solutions to make your projects move forward!
Polyclonal antibody production vs.
monoclonal antibody production
You don’t know how to choose between polyclonal and monoclonal antibodies for your antibody production? Try our free online quiz and get the answer in less than 2 minutes!
When choosing between polyclonal antibodies and monoclonal antibodies, several parameters have to be taken into account. Here is an overview of advantages and drawbacks of each solution.
|Monoclonal antibodies||Polyclonal antibodies|
|Advantages||Better specificity & less background|
Unlimited quantity of antibody
Standardization + repeatability
Possibility to sequence and engineer
|Potentially better immunogenicity + antibody sensitivity|
Less sensitive to antigen’s conformational variations
Fast and inexpensive to produce
|Drawbacks||Potentially lesser immunogenicity + antibody sensitivity|
Long and expensive to produce
|Background risk due to cross-reactions|
Limited quantity of antibody
|Species available||Mouse, rat||Mouse, rat, rabbit, chicken, guinea pig, sheep, goat, alpaca, llama|
Polyclonal antibodies consist in a mixture of antibodies able to bind several epitopes of a same antigen. They are produced by animal immunization and collected directly from serum. The serum is usable as is or can be further purified in order to obtain a serum free solution. Polyclonal antibody production is less complex than monoclonal antibody production. It is therefore the best solution for a fast and inexpensive antibody development. However, polyclonal antibodies are prone to batch-to-batch variations due to the production method.
Polyclonal antibodies present several advantages due to their multiple epitope binding properties:
- they can present high sensitivity and represent a perfect solution for detection of proteins presenting a low expression level
- they are less sensitive to antigen’s conformational variations and can thus be used for denatured protein detection
However, multiple epitope binding leads to some drawbacks as it implies cross-reactivity due to lower specificity. For this reason, polyclonal antibodies are not suitable for quantitative assays or for the recognition of a specific domain. Nevertheless, several methods can limit or even overcome the cross-reactivity issue, as for example:
- immunization with one or several peptide immunogens limiting the recognition of a specific epitope,
- addition of a purification step against the target epitope.
Monoclonal antibodies are generated by identical immune cells which are clones of a single parent cell. Thus, monoclonal antibodies specifically recognize one epitope of an antigen.
First steps of monoclonal antibody generation are similar to polyclonal antibodies. Then, B lymphocytes are isolated and fused with a myeloma cell line leading to an immortalized hybridoma cell line producing the desired monoclonal antibody. Monoclonal antibody production is highly complex and is therefore more expensive than polyclonal antibody production. However, once developed, the hybridoma cell line has the ability to produce an “infinite” quantity of antibodies. Monoclonal antibody production can even be secured by hybridoma sequencing and stable cell line development
Monoclonal antibodies can present several advantages over polyclonal antibodies:
- recognition of a single epitope of an antigen,
- low cross-reactivity,
- high reproducibility,
- possibility to develop large-scale applications (therapeutic antibody development or diagnostic manufacturing).
Due to their high specificity, monoclonal antibodies are more sensitive to epitope’s conformational changes.
Antigen design for antibody production
Our main concern when designing the most appropriate antigen for an antibody production is to use an antigen type that is as close as possible to the target the antibody will finally need to detect in your assay. We try to stick to that rule as much as possible even though some other constraints like specificity for instance may sometimes force us to accept making compromises. Following that rule allows to significantly increase the success rate of developing the perfect antibody for the final application. There are mainly 3 types of antigens which are commonly used for antibody production:
- Proteins: antibody production against a protein remains the best way to make sure that an antibody will recognize the target protein. However, this method increases the risk of cross reaction induced by the recognition of a domain common to several proteins.
- DNA: DNA immunization is commonly used for difficult-to-express proteins or transmembrane proteins. The main advantage of DNA immunization remains the possibility to express the antigen in its native conformation.
- Peptides: using peptides as an antigen favors antibody production with low cross-reactivity and high specificity. However, they are often less immunogenic due to their small size. Designing a peptide antigen for antibody production requires to take several factors into considerations, most of them directly related to the protein structure:
- Avoid peptides including sequences which are common to several protein structures
- Favor flexible and solvent-exposed sequences which will remain accessible for the antibody produced and avoid organized secondary structures such as α-helices or β-sheets
- Favor hydrophilic sequences which are more prone to be solvent-exposed than hydrophobic structure which are generally buried in the structure. They are also easier to solubilize before injection.
Finding the right size for a peptide antigen is not trivial as it is a balance between a sufficient immunogenicity, solubility, specificity, cross-reactivity and native secondary structure. Thus, a sequence of 10-20 amino acids is generally considered as an optimal size for antibody production.
With more than 16 years of experience in antibody production (monoclonal or polyclonal), ProteoGenix guides you through the best antigen design strategy. Please feel free to describe your project to our PhD account managers who will be pleased to bring a real added-value to your antibody production strategy.
How are antibodies produced?
How does it influence the choice of your antibody production technology?
Even if the scope of this page is not to learn how antibodies are produced, understanding the basic steps of antibody production in vivo allows getting a better overview of the advantages of both in vivo and in vitro antibody production. Also, it can partly govern the choice of the right antibody production method.
In vivo antibody production occurs when the immune system encounters a foreign substance, the immunogen. Antibodies are produced by the well-known B cells. Once an antigen binds to the B lymphocyte surface, B cells can be activated by 2 different manners: polysaccharides, lipopolysaccharides, and other non-protein antigens activate B cells in a T cell independent manner (the activation signal comes from another source than T cells such as factors from the complement system) whereas protein antigens activate them in a T cell dependent manner occurring in several steps:
- Antigen recognition and internalization by the B-cells
- Presentation of the antigen to a helper T cell specific to the same antigen
- Helper T cell – antigen interaction (linked recognition)
Once activated by linked recognition, type 2 helper T cells activate B cells thanks to cytokine release leading to proliferation into daughter cells. Activation of B cells leads to somatic hypermutation which results in random mutations of variable heavy and light chains. This step is followed by another selection step in which only the positive mutation (leading to an affinity increasing against the antigen) will be conserved. Thus, somatic hypermutation allows the generation of antibodies and memory cells with higher affinity against the antigen.
Further cytokine secretion results in the differentiation of activated B cells into memory B cells and plasma cells, each having a defined function:
- Memory B cells allows for a fast response in case of a subsequent exposure
- Plasma cells secreting IgMs
Stimulation of plasma cells by the cytokines released by type 2 helper T cells allows switching from IgM production to another class such as IgG, IgA or IgE. This switch does not affect the affinity of the antibodies for the antigen as it only consists in a genetic modification a modification of the constant region (accomplished by genetic rearrangement).
The release of antibodies into the bloodstream occurs only once antibodies with sufficient affinities were generated. This antibody response occurs in two steps:
- Primary response: the primary response is characterized by a latent phase of 10 days approximately. This phase includes all the steps from antigen binding to naïve B cells to clonal proliferation. The end of this period is characterized by an increase of IgM levels in the serum with a maximum IgM at D14. Then, a decrease of IgM level is observed and is correlated with an increase of IgG levels. This phase also include the generation of memory cells which will be activated in case of new exposure.
- Secondary response: during the secondary response, the latent phase does not last more than a few days. Then, this response is characterized by high production levels of high affinity IgG.
In the context of custom antibody generation, the induction of an immune response can be obtained by immunization with various substances such as molecules, viruses or bacteria. However, some substances do not induce an immune response (or only a weak immune response) mostly due to their small size. That’s why, the antigen design remains a major step when producing a custom antibody. While the antigen can be modified to elicit an immune response, it is also possible to administer an adjuvant to the host to obtain a better response. If this is not sufficient, in vivo antibody production might not be adapted to your project and in vitro solutions should be considered. In this case, naive library screening with antibody phage display remains the best solution.
You’re not sure about the best antibody generation technology to use for your project? Contact our antibody production experts who will elaborate a custom solution for your project.