Increase the sensitivity of your immunoassays with our integrated solutions for polyclonal antibody production. From antigen design to serum purification, our antibody production services were designed for maximum quality and production yield. Generate high-quality custom antibodies using any type of antigen and in any species including rabbit, chicken, mouse, rat, goat, sheep, llama, or alpaca!

ANTIGEN Peptide synthesized by ProteoGenix
2 peptides Modified and non-modified peptides
ANTIGEN Recombinant Protein produced by ProteoGenix
ANTIGEN Provided by customer
Immunization of 2 rabbits
Standard (51 days) or Express protocol (28 days)? Standard Standard Standard ExpresswayTM Standard Standard
PURIFICATION vs. Protein A or G
Purification/ depletion
PURIFICATION No purification
Guarantee ELISA titer ELISA titer WB + ELISA titer ELISA titer ELISA titer ELISA titer
Timeline 9-10 weeks ≈15 weeks ≈13-15 weeks 28 days ≈15 weeks ≈15 weeks

Our polyclonal antibody production process

Antigen design

  • Definition of an antigen design strategy for optimized polyclonal antibody production
  • Protein design
  • Peptide design


1-2 days

Designed sequence for customer’s validation

Antigen production


Proteins: 3-5 weeks
Peptides: 3 weeks

Antigen sample (protein or peptide)

Animal immunization

  • Animal injection with the antigen produced or provided + Freund’s adjuvant
    Injection route: subcutaneous, intradermal, intramuscular, intraperitoneal, intravenous


Fast: 28 days
Proteins: 51 days
Peptides: 70 days

Pre-immune serum

Antibody testing

  • Antibody QC analysis : ELISA, Western Blot or Dot Blot (depending on the guarantees)


1-2 days

Final Immune serum (if no purification)

Serum purification

  • Purification against protein A or protein G
  • Purification against antigen
  • No purification


1 week

-Purified polyclonal antibodies (serum of each animal is purified separately)
-Certificate of analysis (CoA)

Antibody conjugation and fragmentation

Our recommendations for polyclonal antibody production

What To Consider When Choosing A Host For Polyclonal Antibody Production

To select the most suitable species for your polyclonal antibody production, several factors need to be taken into account:

Amount Needed

  • Rabbits are the ideal hosts for polyclonal antibody production – they have a convenient size making them easy to handle. Plus, they produce serum with high titers of target-specific antibodies leading to greater antibody production yields. For larger quantities, goats, sheep, llama, or alpaca should be used.

Phylogenetic Distance

  • The greater the phylogenetic distance between the source of the antigen and the host species, the stronger the immune response. For instance, when generating polyclonal antibodies against a highly conserved mammalian antigen, chickens could be a good host species.

Final Application

  • When using polyclonal antibodies in tandem with their monoclonal counterparts, the host species for antibody generation should be phylogenetically distant from each other. For instance, when primary rabbit antibodies are used, secondary/polyclonal antibodies should be generated in species like llama, alpaca, chicken, goat, among others.

Best Reasons To Choose A Host Species For Polyclonal Antibody Generation

Rabbit hosts for polyclonal antibody production


The first choice for polyclonal antibody production given their size, ease of handling, and ability to produce high titers of high-affinity antibodies.

Sheep and goat hosts for polyclonal antibody production

Sheep and Goats

Ideal hosts when larger amounts of antisera are needed.

Chicken hosts for polyclonal antibody production


Ideal hosts when generating antibodies against conserved mammalian proteins. Possibility to harvest antibodies using non-invasive methods.

Llama and alpaca hosts for polyclonal antibody production

Llama and Alpaca

Ideal when targeting cryptic antigens and when the final application requires a higher capacity for tissue penetration and higher antibody stability.

Host selection is an important step of every polyclonal antibody production process. Although rabbit hosts are the most conventional choice, there is a growing interest in producing chicken and camelid antibodies.

Polyclonal antibody production in chicken can be quite advantageous when the process needs to be scaled-up because IgY antibodies are extracted from egg yolk instead of serum. It is known that egg yolk is more challenging to purify than serum; however, it can be produced in higher quantities in comparison to mammalian polyclonal production.

In contrast, camelids are increasingly appreciated as polyclonal antibody production hosts. In addition to conventional IgG antibodies, they can produce immunoglobulins devoid of light chains – heavy chain antibodies (HAbs). These molecules have unique properties including:

  • Increased stability at extreme pH or temperature
  • A low steric hindrance allowing easier access to buried antigens that would not be accessible to conventional antibodies.
  • A higher capacity of tissue penetration is a precious asset for immunohistochemistry experiments or even for therapeutic applications.

Polyclonal antibody production at ProteoGenix: FAQ

What types of antigens can you use for polyclonal antibody production?

We have the capacity to produce all types of antigens at our facilities including peptides, proteins, DNA, small molecules, and cells overexpressing the target antigen. We can also produce polyclonal antibodies using customer-provided antigens.

How to optimize the polyclonal antibody production process?

Antigen Design And Production

Achieving high titers of target-specific antibodies depends on the antigen’s capacity for eliciting a strong immune response. For this reason, choosing a suitable antigen for immunization remains one of the most important steps of the polyclonal antibody production process

Several antigens may be used for polyclonal antibody generation including:

  • Proteins – proteins are the most conventional antigens for antibody generation. They ensure polyclonal antibodies recognize different relevant epitopes naturally exposed in the native conformation of the protein.
  • Peptides – using peptides for immunization is useful when developing antibodies for linear epitopes (important in Western Blot applications) or when developing antibodies against a specific epitope (increases assay specificity). Since most peptides aren’t immunogenic, adjuvants are typically used to enhance the immune response.
  • DNA – genetic immunization is reserved for special cases. For instance, when the target protein is hard to produce, unstable or contains complex transmembrane domains, genetic immunization may be a suitable alternative.

Other antigens may be used for immunization such as small molecules or even whole cells (native or recombinant); however, these projects require the development and testing of custom immunization solutions.

Animal Immunization

Our standard immunization protocol starts at:

  • 51 days for anti-protein polyclonal antibody production
  • 70 days for anti-peptide polyclonal antibody production

Both protocols can be extended if guaranteed antibody titers are not reached. Typically, it is better to immunize animals with a lower quantity of antigen and for longer periods, rather than using higher quantities of antigen and shorter immunization times.


Polyclonal Antibody Purification

Polyclonal antibodies are typically harvested by bleeding the hosts after desired antibody titers are reached. The cellular fraction and the antibody-enriched serum can be separated by centrifugation resulting in a crude polyclonal antibody solution.

Crude preparations are useful for many applications. However, for enhanced sensitivity and reduced off-target binding, these preparations should be purified. Polyclonal antibody purification can be carried out by:

  • Protein A or G purification – these proteins are produced by Staphylococcus aureus and Streptococcus spp., respectively, and they can bind the Fc fragment of antibodies with high affinity. Protein A/G purification allows the straightforward separation of immunoglobulins from all other serum components. However, unspecific antibodies with a low affinity towards the target are not eliminated with this type of purification.
  • Antigen-specific purification – using affinity chromatography to recover polyclonal antibodies with high affinity towards a specific antigen is a widely used process of polyclonal antibody purification. It ensures only the antibodies with the highest affinity are recovered, reducing off-target binding and, consequently, reducing background noise in immunoassays.

Need advice for your custom polyclonal antibody production? Please feel free to contact your dedicated account manager!

How do you measure antibody titers?

To monitor the response to immunization, we recurrently take samples from hyperimmunized hosts to measure their antibody titers. Conventionally, this is done using ELISA, but alternatively, our clients may request antibody titer verification by Western Blot as well.

Polyclonal antibodies

What are polyclonal antibodies?

Polyclonal antibodies are a mixture of monoclonal antibodies that originated in different B cell clones. Consequently, these antibodies display different epitope-specificity and binding affinity. This property makes them highly sensitive to low abundance markers and highly effective at tackling complex targets.

How are polyclonal antibodies classified?

Given their diverse nature, polyclonal antibodies are often classified according to the host species and antigen used for their generation. These antibodies are generally produced in rabbits, goats, and sheep. Rabbit polyclonal antibodies are particularly useful because the rabbit’s immune system can generate high-affinity antibodies in higher abundance than other host species.
Polyclonal antibodies are also named according to this classification, in this way, anti-mouse rabbit polyclonal antibodies are pAbs designed to bind to murine IgG and produced in a rabbit host.

What are the main advantages and uses of polyclonal antibodies?

Polyclonal antibodies are a mixture of monoclonal antibodies with distinct epitope-specificity and binding affinity. They are produced by immunizing animal hosts with a specific target (protein, peptide, DNA, etc.) and harvested by separation and purification of their serum after the development of a strong immune response.

Unlike monoclonal antibodies, prized for their high specificity and selectivity, polyclonal antibodies are known for their enhanced sensitivity. This property makes them invaluable tools for multiple applications including research (basic, medical, etc.), therapy, and diagnostics. They are particularly advantageous reagents to detect low abundance markers, toxins, among other substances. The detection of these rare markers is extremely useful for the design of early diagnostic tools, food monitoring applications, and for capturing rare targets and purifying them before further analysis.

From a therapeutic perspective, polyclonal antisera are invaluable to treat complex acute conditions like snakebite envenoming. Venoms cause acute reactions and are composed of a complex mixture of different proteins with key roles in pathogenesis. In this way, successful antivenom therapies need to be able to tackle several antigens and epitopes present in a single venom to block the pathogenic pathways and avoid adverse reactions.

From a technical point of view, polyclonal antibodies are cheaper to produce than their monoclonal counterparts. Plus, the timelines for polyclonal antibody production are significantly shorter than the lead time of monoclonal antibody production. In contrast, their most notable limitations are the difficulty in scaling up the process and high batch-to-batch variability. The impact of the latter can be mitigated by including proper controls (positive and negative) and standards in every assay.

Overcoming scale-up hurdles is more challenging, but recent breakthroughs have provided interesting solutions. One of the most interesting solutions is the purification of IgY antibodies present in egg yolk using ionic liquids. Due to the non-invasive methods of antibody harvesting and the cost-effective new purification methods, these molecules may soon gain ground over other polyclonal antibodies for a multitude of applications.

What are the advantages of pairing monoclonal and polyclonal antibodies?

Polyclonal antibodies can be used by themselves or in tandem with their monoclonal counterparts in different assays. The most common application of polyclonal and monoclonal antibody pairs is the enzyme-linked immunosorbent assay (ELISA). In ELISA, monoclonal antibodies (also called primary antibodies) are typically used to bind a specific target, while polyclonal antibodies (secondary antibodies) are used to bind the primary antibodies.

In this configuration, the primary antibodies are “naked” (no tags) while the secondary antibodies carry the enzymatic tag. This format allows the amplification of the detection signal, lowering detection thresholds considerably. The signal amplification occurs because, while primary antibody binds to a single epitope on the target molecule, secondary antibodies bind to multiple regions of the primary antibodies. In this way, multiple secondary antibodies conjugated with an antibody can coat the surface of the primary antibody, dramatically enhancing the detection signal.

The enhanced sensitivity of these molecules comes with its own set of specific hurdles. For instance, polyclonal antibodies are notorious for their propensity for off-target binding, which may lead to false-positive results. To overcome this hurdle, polyclonal antibodies must be submitted to stricter purification processes (e.g. by performing antigen-specific antibody purification). Additionally, the inclusion of controls and standards can help researchers correctly identify thresholds of detection.

Polyclonal antibody production:
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