Polyclonal Antibodies

Custom Polyclonal Antibody Production

All APS antibody projects are produced in the UK under Home Office licence. This ensures the highest quality of care for the host animals and the production of premier quality antibodies.

Overview

The APS Custom Polyclonal Project unit holds a licence under the Animals (Scientific Procedures) Act. Our facilities are regulated and controlled under strict UK Home Office guidelines ensuring a high standard of animal care by experienced and dedicated staff. Further information may be found on our Project Requirements section of the INFORMATION page.

Download APS Custom Polyclonal Antibody Production pdf document

Custom Polyclonal Projects

These projects typically follow the following structure:

 

Step One

Antigens

The immunogenicity of the chosen antigen is of paramount importance to the success of any custom polyclonal antibody project. The choice of antigen is discussed on the PDF document: Antigen Selection and Preparation. Once the antigen has been chosen, typical recommended antigen concentration per immunisation are as follows:


If you have any queries concerning the choice of antigen or buffer please contact our technical team for advice.

Adjuvant

The adjuvant Complete Freunds is typically used for primary immunisation, with subsequent immunisations using Incomplete Freunds.

In order to eliminate or reduce discomfort APS pre-screens batches of adjuvant and reserves suitable batches.

A number of other adjuvants are available on request.

 

Step Two

Host Animals

APS can offer one of the widest choice of animals for polyclonal antibody production in the UK. The most suitable host for antibody production will depend on the amount of antibody needed and the characteristics of the antibodies being made.

 

Rabbits

Rabbits are suitable for most projects. They produce strong immune reactions against most antigens in a relatively short period of time. The raw anti-sera can be used directly in assays. Rabbits typically give between 40ml and 90ml of serum.

Sheep & Goats

Sheep and goats give strong immune responses against most antigens and produce large volumes of serum, typically 200ml-300ml per host. They are the host of choice if large volumes of antisera is required. Sheep and goats can be kept on long term project and bleeds can be taken monthly for a period of 5 consecutive months. The average serum yield per annum is 2500ml.

Chickens

Chickens (eggs) should be considered as the host if it is important for the phylogenetic relationship between the antigen donor and the antibody producer to be distant. Chickens transfer high quantities of immunoglobulins, (IgY), into the egg yolk, which eliminates the need for invasive bleeding procedures. The resultant antibody will need to be purified before use.

Mice & Rats

Mice and rats are ideally used as hosts for polyclonal projects when screening potential antigens prior to immunisation into a larger host. These hosts only provide small amounts of whole antisera.

Alpaca & Llama

Alpaca and llama are unique in producing antibodies composed solely of two identical heavy chains which makes them of great interest for basic research and ther apeutic applications. Alpaca antibodies are robust, with the ability to withstand elevated temperatures and a wide pH range, extending their within in vivo and in vitro applications.

Emu

Emu (eggs) offer an alternative to the chicken egg for the production of IgY. An emu egg is typically 10 -15 times larger than a chicken egg.
NOTE: Availability is seasonal, from October to March.

 

Step Three

Immunisation Protocols

Protocols

Once the most suitable host has been chosen for a custom polyclonal project it is necessary to consider the design of the immunisation protocol. Not all host species elicit the same immune response to a specific antigen. APS has developed a series of host specific protocols enabling the optimum immune response over a specific period of time. These protocols may be modified to suit individual circumstances following discussion between the end user and the technical team. Full details of the immunisation schedule are supplied to the customer prior to the commencement of the project. Any changes in the protocol are carried out only with the agreement of the customer following discussions with the technical team.

In order to monitor the effectiveness of the protocol being followed and the immunogenicity of the antigen, test bleeds are taken at regular, predetermined intervals. Following monitoring of these test bleeds by ELISA the immunisation protocol may be modified, if required.

 

ELISA

ELISA monitoring of test bleeds during the progress of a custom polyclonal project is a simple and convenient way to follow the development of the immunoreactivity in each host during the immunisation protocol. During this process, the test sera is coated onto adapted microtitre plates and several parameters are reviewed including:

 

ELISA screening may be carried out by APS or then the test bleed samples together with a pre-immune sample bleed from each host may be returned to the customer in order that screening may be carried out in their own facility.

 

Step Four

Project Termination

Following the successful completion of the custom polyclonal project protocol the terminal bleeds, processed into serum, are supplied to the customer in entirety. The resulting serum will contain the whole antibody as well as other protein types. Whole antisera may be suitable for use in many applications, but it may be necessary to improve specificity by applying purification steps to the whole serum.

Please see our Purification section on our SERVICES page for further options, or please contact our technical team for further information.

 

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