Polyclonal Antibody Generation

BioGenes is your expert for high-quality polyclonal antibody development services. Our dedicated team has developed more than 10,000 polyclonal antibodies for more than 30 years. Our immunization protocols deliver fast results, high yields, and uphold superior quality. All animals used for polyclonal antibody generation are housed in certified facilities in Germany demonstrating our commitment and responsibility for animal welfare. Below, you'll find detailed protocols for immunizing rabbits and goats. Should you require immunization of rats, mice or sheep, please don't hesitate to contact us.

 

Development Strategy

Tailored to meet your exact requirements, our polyclonal antibody development services offer customization across every stage of the process: From initial consultation to antibody generation, purification, and stringent quality control of purified antibodies, we ensure a seamless workflow. Our services cater to diverse applications, e.g. for the development of an ELISA.

Phase 1 - Expert consultation

  • Selection of species and immunization protocol

Phase 2 - Preparation of the immunogen

Phase 3 - Immunization

  • According to the selected species and protocol

Phase 4 - ELISA titer tests

  • Of pre-immune sera and antisera 

Phase 5 - QC & deliverables

  • Delivery of antiserum
  • opt.: Immunoglobulin-specific or antigen-specific purification of antibodies
  • opt.: Dialysis of purified antibodies to PBS or other desired buffers
  • opt.: Quality control and characterization of purified antibodies using SDS-Page, 1D Wesern Blot or ELISA

Approximate project duration: 5 weeks

Extended protocols available.

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Immunization Protocols

Protein Immunization of Rabbits

We offer various immunization protocols for generating polyclonal antibodies in rabbits, which can be adapted in consultation with our scientists. Assuming the antigen is suitable, the immune response will be controlled by ELISA. After delivery of the antiserum, the animals can be kept for further antiserum production.

Prior to the start of the project, the customer can decide whether the sera shall be delivered after each bleeding day or collectively after completion of the immunization protocol. Before administering the 1st immunization, every protocol includes the delivery of 1.5 mL pre-immune serum. If desired, we can provide assistance in selecting suitable animals for immunization and conduct pre-immune serum testing by ELISA.

Our shortest protocol spans 28 days. However, in instances where the antigen exhibits diminutive size or limited immunogenicity, we extend the protocol to 42 days or longer to ensure a robust immune response and adequate antibody production. For enhanced quality control, we provide repetitive titer measurements. 

In case a higher amount of antiserum is desired, we integrate booster injections into the protocol regimen. While this prolongs the duration, it results in a higher yield of antiserum by the end of the process. Additionally, we offer a „final bleed” option at the end of the protocol to maximize the volume of antiserum collected.

 

ProtocolBleedsRequired antigen amount* (per 2 rabbits)      Deliverable antiserum (per rabbit)ELISA titer measurementOptional:final bleeding  
28 days11.7 mg15  mLat day 28x
42 days11.7 mg15  mLat day 42x
42 days21.9 mg30  mLat day 2850 mL
91 days32.2 mg45  mLat day 4250 mL
105 days42.7 mg60  mLat day 42, 63, 84, 10550 mL

* Kindly be informed that BioGenes does not accept genetically modified organisms (GMOs) for rabbit immunization. Safety level 1 (S1) and safety level 2 (S2) requirements are applicable only to the laboratories handling mice.

 

Peptide Immunization of Rabbits

At BioGenes, we offer specialized protocols for the immunization of rabbits with peptides. These protocols were carefully designed with a focus on the peptide as the antigen, aiming to achieve maximum antibody affinity. In case of small molecules, such as peptides, haptens, or proteins below 12 kDa, conjugation to a carrier protein becomes necessary. The choice of carrier is typically discussed and agreed upon with the customer. Hemocyanin is commonly used for immunization, while a protein carrier such as bovine serum albumin (BSA), not utilized during immunization, is employed for titer determination. This strategic approach ensures the detection of peptide-specific antibody titers. 

Our scientists will assist you in the selection of the sequence and conjugation to a carrier protein. We recommend a sequence of 15 amino acids, which will be synthesized and purified by our collaborators.

 

ProtocolBleedsDeliverable antiserum (per rabbit)ELISA titer measurementOptional: final bleeding
42 days115 mLat day 42x
56 days230 mLat day 4250 mL
91 days345 mLat day 4250 mL
105 days460 mLat day 42, 63, 84, 10550 mL

 

Protein Immunization of Goats

Immunization of goats leads to large amounts of antiserum and/or purified antibodies with manageable time frames and animal numbers. The immunization schedules can be modified according to your special requirements and the aim of your project. We conduct a quality control of the antiserum using ELISA, if the antigen is suitable. Animals will be kept after delivery of the antiserum.

 

ProtocolBleedsRequired antigen amount* (per 2 goats)      Deliverable antiserum (per goat)ELISA titer measurementOptional:final bleeding  
42 days115 mg100  mLat day 42x
105 days421 mg400  mLat day 42, 63, 84, 105300-500 mL

Additional Services & Information

Below you can find additional information regarding services, resources and antigen requirements.

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If you need more information, a specific offer 
or want to talk to an expert

FAQs

What distinguishes polyclonal antibodies from monoclonal antibodies?

Polyclonal antibodies are composed of a mixture of antibodies produced by different B cell clones. They exhibit varied specificity, selectivity, and antigen-binding affinity, recognizing different epitopes of a target molecule.

Which animals are immunized?

BioGenes utilizes a variety of host species for polyclonal antibody development, including rabbits, mice, rats, sheep, and goats.

What are the main applications of polyclonal antibodies?

Polyclonal antibodies are utilized in immunoassays such as immunohistochemistry, ELISA, Western Blot, flow cytometry, immunoprecipitation, as well as in immunotherapy for diseases like cancer and infectious diseases.

Why are polyclonal antibodies considered valuable in various applications?

Polyclonal antibodies offer multi-epitope specificity, making them highly sensitive and effective in detecting or capturing low-abundance targets. They are utilized in diverse applications, including research and diagnostics, due to their versatility.

How do polyclonal antibodies contribute to early detection in cancer and autoimmune markers?

Due to their enhanced sensitivity, polyclonal antibodies are crucial for early detection of markers related to cancer, autoimmunity, and other conditions. This sensitivity is essential for preventing unfavorable disease outcomes.

What challenges are associated with polyclonal antibodies?

Challenges include variability in production yields based on the selected species, animal size and immunization method. Additionally, the variability in antigen selectivity, specificity, and binding affinity necessitates careful consideration in immunoassays, requiring proper control samples.

Can you provide guidance on selecting the most suitable immunization protocol?

Our experienced team can assist you in choosing the right immunization protocol tailored to your objectives. Factors to consider include the type of antigen (protein or peptide), desired antibody characteristics, and the timeframe for antibody development.

How are polyclonal antibodies purified?

Polyclonal antibodies are purified using either immunoglobulin-specific or antigen-specific methods. It's worth noting that the ultimate decision regarding purification methods typically rests with the customer.

  • Immunoglobulin-Specific Purification:
    • Protein A or protein G purification: Protein A or protein G is used to efficiently purify polyclonal antibodies, particularly IgG immunoglobulins. These proteins interact with the Fc region, allowing straightforward recovery from crude antisera.
  • Antigen-Specific Purification:
    • Using the target antigen: Antigen-specific purification involves using the target antigen to further purify polyclonal antibodies.

How long can animals be kept?

The duration of animal housing is flexible and can be tailored to meet the specific protocol requirements of the project.

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