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 polyclonal antibodies using any type of antigen and in any species including rabbit, chicken, mouse, rat, goat, sheep, llama, or alpaca!
Why choose ProteoGenix’ polyclonal
antibody production services?
All our packages include a high antibody titer guarantee (1/64000 in ELISA). Our anti-protein package even includes WB positive results.
With all-included packages starting from 255€, we provide the most competitive price on the market!
Instant online order
Save time and buy polyclonal antibodies directly online thanks to our online form!
Large choice of species
Choose between different species depending on the volume and format of antibodies you require!
We offer integrated polyclonal antibody production solutions from antigen design to purification and conjugation (if requested).
Get your polyclonal antibodies in 28 days thanks to our ExpresswayTM protocol.
Our polyclonal antibody production packages
|ANTIGEN||Peptide synthesized by ProteoGenix||2 peptides||Modified and non-modified peptides|
|Recombinant Protein produced by ProteoGenix|
|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|
|vs. Antigen||Purification/ depletion|
|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
- Definition of an antigen design strategy for optimized polyclonal antibody production
- Protein design
- Peptide design
- Protein production in an expression system of your choice
- Peptide synthesisand conjugation to a carrier (KLH, BSA or OVA) to improve immune response
- Animal injection with the antigen produced or provided + Freund’s adjuvant
Injection route: subcutaneous, intradermal, intramuscular, intraperitoneal, intravenous
- Antibody QC analysis : ELISA, Western Blot or Dot Blot (depending on the guarantees)
- Purification against protein A or protein G
- Purification against antigen
- No purification
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:
- 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.
- 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.
- 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
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 Goats
Ideal hosts when larger amounts of antisera are needed.
Ideal hosts when generating antibodies against conserved mammalian proteins. Possibility to harvest antibodies using non-invasive methods.
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.
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.
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.
Pressed for time in your polyclonal antibody production project?
Our exclusive protocol – ExpresswayTM – was designed to generate high-quality polyclonal antibodies in only 28 days.
This is made possible thanks to our special adjuvant aimed at enhancing the immune response.
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.
For more answers to the most frequently asked questions about polyclonal antibodies, consult our dedicated page.
Need advice for your custom polyclonal antibody production? Please feel free to contact your dedicated account manager!