Engaged in the development of immunotherapies or immunodiagnostics? Get custom-made antibodies with optimal properties and exceptional sensitivity, affinity, and stability. Drawing from 25+ years of experience and 6,000+ successful projects, we aim at matching your most innovative and ambitious projects with our most skilled experts and cutting-edge solutions. Whether you are developing monoclonal or polyclonal antibodies to use in therapy, research, or diagnostics, our vast capabilities, and state-of-the-art equipment are designed to enable your groundbreaking discoveries.

Not sure which antibody production process is more adapted to your needs? Get a clear and personalized answer in less than two minutes thanks to our online quiz:

Our access to state-of-the-art technologies and immense flexibility helps us to create unique solutions tailored to your specific goals and needs. All our resources and expertise are placed at your disposal to empower your groundbreaking discoveries in antibody development.

Discover how ProteoGenix can help you bring your projects to life.

Our antibody production process

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 are currently developing a new interactive form to guide you to define the most relevant antibody productionstrategy for your project.

Antigen Design Production

  • Broad antigen diversity:
    Proteins, peptides, DNA, small molecules, whole cells

Antibody Generation

3 discovery technologies:

  • Hybridoma generation
  • Naïve antibody phage display (premium libraries adapted to therapeutic and analytical applications)
  • Immune antibody phage display (optimized library construction protocols)

Antibody Characterization & Validation

  • Extensive analytic capabilities for optimal developability: Antibody sequencing, purity, stability, affinity, aggregation, glycosylation profile, and validation in target application (research and diagnostics) or using your antigen’s native conformation (therapeutic development)

Antibody Engineering

  • Diversified strategies for antibody lead optimization: Antibody humanization, affinity maturation, bispecific antibody development, antibody reformatting, antibody-drug conjugation, antibody conjugates for immunoassay development

Antibody Production

Optimized antibody production technologies:

  • Small and medium-scale : transient production in mammalian cells (XtenCHO™, CHO, HEK)
  • Large-scale: stable cell line development in CHO or HEK cell lines
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 Batch variability
Species available Mouse, rat, llama, alpaca, rabbit, human Mouse, rat, rabbit, chicken, guinea pig, sheep, goat, alpaca, llama

Polyclonal antibodies consist of a mixture of monoclonal antibodies able to bind different epitopes of the same antigen. The polyclonal antibody production process starts by immunizing animal hosts, gauging their immune responses, and collecting the antibody-enriched serum when desired antibody titers are reached.

The serum is usable as is or can be further purified. Different purification techniques can be used depending on the final application. For instance, most polyclonal antibodies are purified using immunoglobulin-binding proteins (A, G, or L). However, using this method, many unspecific antibodies are still present in the final production batch. For applications requiring a higher degree of specificity, the serum needs to be purified by antigen-specific methods.

Overall, polyclonal antibody production is less complex than monoclonal antibody production. It is therefore the best solution for fast and inexpensive antibody development. These reagents present several advantages due to their multi-epitope binding properties:

  • High sensitivity – perfect solution for detecting proteins with low expression levels
  • Less sensitive to conformational changes – less sensitivity to antigen’s conformational variations makes polyclonal antibodies useful for denatured protein detection

However, these reagents are prone to batch-to-batch variations due to the antibody production processes used for their generation. Plus, their multi-epitope binding ability may results in an increased cross-reactivity potentially leading to inconclusive findings. Several methods can limit or even overcome the cross-reactivity issue:

  • Immunizing with one or several immunogenic peptides thus limiting the range of epitopes bound by the polyclonal antibody preparation
  • Performing epitope (or multi-epitope) specific purification to limit the regions of the antigen recognized by the polyclonal antibodies

Polyclonal antibodies are prized for their sensitivity and broad reactivity. However, given their batch-to-batch variability, they are not suitable for quantitative assays or assays requiring high specificity.

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.

Unlike polyclonal molecules, monoclonal antibodies can be generated by a variety of methods either using immunization-dependent or independent approaches. The two most important methods of monoclonal antibody production include:

  1. Hybridoma development ( in vivo )
  2. Antibody phage display ( in vitro )

When generated in vivo, the process of custom monoclonal antibody generation is very similar to the process of polyclonal antibody production. But once an effective immune response is mounted, plasma cells are isolated from the serum and fused with myeloma cell lines to produce hybridomas – immortalized plasma cells.

Despite the complexity of this process, once developed, hybridomas can produce an “infinite” quantity of antibodies. Plus, hybridoma cell lines can be sequenced and quickly adapted to recombinant production – which would ensure the long-lasting and large-scale production of these antibodies.

The process of in vitro antibody generation (phage display) is often favored for therapeutic applications. When screening vast libraries of naïve or immune antibodies, the probability of finding antibodies able to bind novel targets is immense.

Overall, monoclonal antibodies can present several advantages over polyclonal antibodies:

  • High specificity: ability to recognize a single epitope of an antigen and detect specific post-translational modifications or single-nucleotide mutations
  • High selectivity: monoclonal antibodies typically have high selectivity (low cross-reactivity), and thus have lower risks of off-target binding
  • High batch-to-batch consistency: allowing standardization of antibody-based assays and the quantification of targets
  • Scalability: possibility of developing large-scale applications (therapeutic antibody development or in vitro diagnostic or medical devices manufacturing).

Due to their high specificity, monoclonal antibodies are more sensitive to epitope’s conformational changes. This property makes them less effective in applications that require higher sensitivity (low-abundance targets) but it also makes them advantageous for applications requiring a high selective such as differential diagnostics (detection of mutations or post-translational modifications) or treatment of complex diseases.

Why is proper antigen design so crucial
the production of high-quality antibodies?

Drawing from our extensive experience in antibody development, we know antigen design can make or break a project. To maximize your chances of success, we always consider the native conformation of your antigen and the final intended application of your antibody.

For this reason, we continuously invest in antigen design diversification:

  • Proteins: these classic antigens remain the most important approach to antibody generation. The use of whole recombinant proteins ensures your antibodies recognize a single antigen (high specificity). However, this method also increases the risks of generating antibodies with low selectivity and prone to binding conserved domains shared by many different proteins. At ProteoGenix, we can produce proteins in a variety of recombinant systems, including:

Mammalian cells

Produce your complex protein antigens in CHO or HEK cell lines – the best mammalian hosts on the market

Bacterial cells

Generate your antigens in Escherichia coli or Bacillus subtilis for high productivity


Learn more about our protein production protocols in Saccharomyces cerevisiae or Pichia pastoris

Insect cells

Produce your protein antigens in our baculovirus/insect cells for improved protein folding

  • 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 taking several factors into considerations:
    • Avoid peptides containing sequences that are conserved among several protein families
    • Favor flexible and solvent-exposed sequences (no secondary structures such as α-helices or β-sheets) which will remain accessible throughout the process of antibody generation
    • Favor hydrophilic sequences which are more prone to be solvent-exposed than hydrophobic structures, generally buried in the structure. They are also easier to solubilize before injection.
    • Favor peptides with 10-20 amino acids (optimal size) which offer the most balance between immunogenicity, solubility, specificity, cross-reactivity
  • Other antigens: whole cells, small molecules, etc.

Our track record of 6,000 antibodies produced (monoclonal or polyclonal), allows you to make empowered decisions regarding antigen design. Please feel free to describe your project to our PhD account managers who will be pleased to bring real added-value to your antibody production strategy.

How are antibodies produced?
How does it influence the choice of your antibody production technology?

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 methods. Also, the particular advantages of each method may dictate the best choice for your projects.

In vivo antibody production occurs when the immune system encounters a foreign substance, the immunogen. Once an antigen binds to the surface of a B lymphocyte, these cells can be activated in 2 different ways: polysaccharides, lipopolysaccharides, and other non-protein antigens activate B cells directly 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 the proliferation of daughter cells. This process culminates in 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 increased affinity 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 are non-secreting and instead act as the immunogenic memory allowing a fast response in case of subsequent exposure to the same antigen
  • Plasma cells secreting IgMs that subsequently undergo isotype switching to produce IgG, IgA, and IgE antibodies

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 are 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 in IgM levels is observed and is correlated with an increase in IgG levels. This phase also includes 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.
Primary and secondary antibody response

Adapted from: Abbas et al. Cellular and Molecular Immunology. Elsevier.


In the context of custom antibody generation, eliciting an immune response can be achieved by immunizing with various substances such as large 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.

For this reason, antigen design remains a major step when producing a custom antibody. While the antigen itself can be modified to elicit an immune response, it is also possible to co-administer it with an adjuvant to enhance its immunogenicity. If chemical adjuvants are insufficient, 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&lt remains the best solution.

Not sure about the best antibody generation technology to use for your project? Contact our antibody production experts who will elaborate a personalized solution adapted to your needs, budget, and time.