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. We offer a variety of host species, and our immunization protocols ensure fast results, high yields, and a superior quality.

 

Development Strategy

Every polyclonal antibody development can be customized according to your specific needs and can cover any or all of the workflow steps — consultation, peptide or protein antigen production, antibody generation and purification – for a variety of 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

  • Of pre-immune sera and antisera, optional affinity purified antibodies

In addition, we are pleased to provide the following services for rabbits, goats and sheep:

  • Immunoglobulin-Specific Purification: Protein A or protein G purification to efficiently isolate polyclonal antibodies, particularly IgG immunoglobulins.
  • Antigen-Specific Purification: Employing the target antigen to further refine the purity of polyclonal antibodies.
  • Dialysis Services: Offering dialysis of purified antibodies to PBS or other desired buffers.
  • Quality Control: Conducting SDS-PAGE analysis under reducing and non-reducing conditions to ensure the purity and functionality of the purified antibodies.
  • 1D Western Blot to characterize the purified antibodies
  • ELISA to characterize the purified antibodies

Approximate project duration: 5 weeks

Extended protocols available.

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

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Animals and Protocols

 

All animals used for polyclonal antibody generation are housed in certified facilities in Germany demonstrating our commitment and responsibility for animal welfare.

Rabbits

Protein Immunization Protocols

At BioGenes, we offer various immunization protocols for generating polyclonal antibodies in rabbits, tailored to meet specific needs such as rapid production, high specificity, or large-scale quantities. The length of our protocols varies depending on the antigen and the desired outcomes. Certain protocols 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.

To obtain an overview of the protocols, timeframes, and bleed schedules, please refer to the following table:

 

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 Protocols

At BioGenes, we offer specialized protocols for the immunization of rabbits with peptides. These protocols are meticulously 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. This process ensures efficient immunization and accurate titer determination.

Peptides and modified haptens must be bound to distinct carrier proteins using different linkers for immunization and titer determination. 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.

To obtain an overview of the protocols, timeframes, and bleed schedules, please refer to the following table:

 

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

Mice

Immunization of mice is a cost-effective way to generate polyclonal antibodies if only small amounts of antiserum are required. We conduct a quality control of the antiserum using ELISA, if antigen is suitable. Immunization of mice with peptide antigens is possible upon request

 

ProtocolAntigen provided by the customerProducts sent to the customerExpected time frame
Standard0.5 mg (≈ 1 mg/mL)0.2 - 0.4 mL antiserum per animal42 days

Rats

Immunization of rats leads to higher amount of antiserum compared with mice and is still fast and cost effective. We conduct a quality control of the antiserum using ELISA, if antigen is suitable. Immunization of rats with peptide antigens is possible upon request

 

ProtocolAntigen provided by the customerProducts sent to the customerExpected time frame
Standard0.5 mg (≈ 1 mg/mL)2 - 4 mL antiserum per animal42 days

Sheep & Goats

Immunization of goats or sheep lead 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.

 

ProtocolRequired antigen amount Deliverable antiserum 
42 days11 mg protein (≈ 5 mg/mL)Pre-immune serum and 100 mL antiserum per animal
105 days21 mg protein (≈ 5 mg/mL)Pre-immune serum and 400 mL antiserum per animal

Additional Services & Information

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

Contact 

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.

How do I determine which host species to choose for my polyclonal antibody development project?

The choice of host species depends on your project's specific requirements. Rabbits, mice, rats, sheep, and goats are available options. Our experts can help you make an informed decision based on factors such as antibody yield, project timeline, and antigen compatibility.

What are the key considerations when deciding between rabbit and mouse as host species?

Rabbits are suitable for larger quantities of antiserum, making them ideal for projects requiring high yields. Mice, on the other hand, are cost-effective for smaller volumes of antiserum. The choice depends on your project's scale and budget.

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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