Polyclonal Antibody Discovery Services

Overview Significance Workflows Key Features Host Species FAQ Case study

Overview of Polyclonal Antibody (pAb)

pAbs are characterized as a collection of monoclonal antibodies (mAbs) capable of targeting specific epitopes. The generation of pAbs is a complex procedure. First, the host is immunized with specific targets, such as DNA, peptides, proteins, followed by separation of serum after a strong immune response, and finally the desired pAbs are harvested by purifying the serum. Typically, pAbs are categorized according to host species and antigens, for example, anti-rat rabbit pAb is designed to conjugate to rat IgG and is raised in a rabbit host.

Why Develop pAbs?

The dominant feature of pAbs is the enhanced sensitivity, a property that allows them to become promising tools for scientific research and diagnosis. The data show that pAbs can effectively detect rare markers. The detection of these low-abundance markers offers great promise for the design of food monitoring tools as well as early diagnostic tools. It is noteworthy that polyclonal antisera plays a crucial role in the treatment of complex acute conditions such as snake bite envenoming, with the property of blocking pathogenic pathways and avoiding adverse reactions.

Fig. 1 A workflow of polyclonal antibody discovery, which includes antigen preparation, immunization, and purification. (Seixas, et al., 2022) Fig. 1 Summary of polyclonal IgY production process.¹

Workflow for Producing pAbs

Procedures	Duration	Description
Antigen Design	2 Days	Design polypeptide or protein sequences according to specific requirements.
Antigen Production	12-30 Days	Produce your target peptide or protein antigen in your desired expression system.
Animal Immunization	45 Days	Animal injection or provided antigen + Freund's adjuvant injection to produce pre-immune serum.
Titration	2 Days	Various assays are utilized for antibody QC analysis such as SDS-PAGE, Western Blot, and ELISA.
Serum Purification	6-12 Days	Serum or egg purification to obtain the final delivered pAb product.
Antibody QC	6 Days	Antibody evaluation by ELISA and WB or other assay required.

Key Features

Various host options

Allowing customers to select the suitable host from multiple options based on the desired antibody characteristics.

Optimal immunization

Our validated antigen strategies significantly improve antibody affinity and specificity.

Flexibility

We are capable of scaling up your project to quickly obtain the final antibody in the quantities you need.

One-stop solution

Our pAb team aims to provide pAb production solutions from antigen design to final binding assay.

Hosts Available at Creative Biolabs

Rabbits

The most popular host that is suitable for antigen immunization. Rabbits are medium-sized, easy to handle, and can directly perform initial analysis of serum.

Chicken

Chicken is suitable for the production of antibodies against mammalian proteins, and these hosts allow for the acquisition of target antibodies in a non-invasive manner.

Sheep and Goats

As large mammals, sheep and goats are ideal hosts for obtaining larger quantities of antisera.

Rats

Rat only requires small amounts of antigen and allows for direct initial analysis of sera. They are suitable for small-scale production of polyclonal antibodies.

FAQ

What are the characteristics of pAbs compared to mAbs?
pAbs are stable across a wide range of pH values and buffer compositions and can detect proteins with high homology to the immunogen. From a technical perspective, pAbs have lower production costs and shorter production cycles.
What factors need to be considered when choosing a host?
To select the most appropriate species for polyclonal antibody production, several factors need to be considered, including the amount of antibody required, phylogenetic distance, and intended application.
Why phylogenetic distance is a key factor in host selection?
In general, the phylogenetic distance between the antigen source and the host species is positively correlated with the strength of the immune response. Therefore, chickens are considered to be an ideal host for the production of pAbs against mammalian antigens.

Case Study

Case 1 - Chicken Polyclonal Antibody Discovery

Fig. 2 A HPLC data suggests the prepared antigen is of good quality. (Creative Biolabs Original) Fig. 2 HPLC analysis of target protein.

Fig. 3 A pre-immune sera testing results to ensure the host chicken does not have a natural immune response against the antigen. (Creative Biolabs Original) Fig. 3 Titration ELISA of pre-immune.

Fig. 4 A 2nd titration result confirms immune response has been properly triggered by the host chicken. (Creative Biolabs Original) Fig. 4 2^nd titration ELISA.

Fig. 5 An electrophoresis analysis to ensure the polyclonal antibodies have been purified successfully. (Creative Biolabs Original) Fig. 5 Gel electrophoresis analysis.

Case 2 - Rabbit Polyclonal Antibody Discovery

Fig. 6 Another 1st titration result confirms immune response has also been properly triggered by the host chicken in case 2. (Creative Biolabs Original) Fig. 6 1^st titration ELISA.

Fig. 7 A 2nd titration result confirms the immune response becomes better than the 1st titration. (Creative Biolabs Original) Fig. 7 2^nd titration ELISA.

Fig. 8 A SDS-PAGE result confirms the polyclonal antibodies have been purified in high purity. (Creative Biolabs Original) Fig. 8 SDS-PAGE of purified pAb.

Fig. 9 A QC assay ensures the polyclonal antibodies can recognize the target properly. (Creative Biolabs Original) Fig. 9 QC of purified pAb by ELISA.

At Creative Biolabs, our stringent quality control gives our customers confidence in the pAbs delivered. Contact us to discuss your unique solution.

Reference

Seixas, António MM, Sílvia A. Sousa, and Jorge H. Leitão. "Antibody-based immunotherapies as a tool for tackling multidrug-resistant bacterial infections." Vaccines 10.11 (2022): 1789. Distributed under Open Access license CC BY 4.0, without modification.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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