Polyclonal Antibody Development

The immune response to an antigen generally involves the activation of multiple B-lymphocytes all of which target a specific epitope on the antigen in question. As a result a large number of antibodies are produced with different specificities and epitope affinities these are known as polyclonal antibodies. 

The immune system can be manipulated to produce large quantities of these polyclonal antibodies for use in a wide range of applications including assay development.  For production purposes these antibodies are generally purified from the serum of immunised animals, were an antigen or immunogen has been injected to stimulate the B-cells to produce a diverse range of immunoglobulin’s specific to that antigen.

 A wide range of naturally occurring and synthetic compounds can be used as immunogens during the antibody production process. In most cases the immunogen will be conjugated to a carrier protein such as bovine serum albumin (BSA) and administered along with an adjuvant to ensure optimum immune response.  The levels of antibody produced increases with repeated exposure to the immunogen of interest as such the animal is injected or immunised at regular time points to achieve the required amount of high affinity antibodies. 

Randox Life Sciences have considerable experience and expertise in custom polyclonal antibody development.  Customised antibodies to a wide range of targets are available from 10-12 weeks after primary immunisation, although prolonged immunisation may be necessary to meet the required specifications. Blood samples are obtained from the sheep throughout the production process to measure the amount of antibody produced.  Once a significantly high titre has been reached the antiserum is prepared by blood collection followed by serum preparation and subsequent purification.

The following is an example of our polyclonal antibody production protocol in Sheep:

Time

Action

 

Serum/Sheep

0

Primary Immunisation

Freunds Complete Adjuvant (CFA)

-

1 Month

Immunisation Boost 1

Freunds Incomplete Adjuvant (IFA)

 

1 Month + ~10days

Production Bleed 1

 

~200mls

2 Months

Immunisation Boost 2

Freunds Incomplete Adjuvant (IFA)

 

2 Months + ~10days

Production Bleed 2

 

~200mls

3 Months

Immunisation Boost 3

Freunds Incomplete Adjuvant (IFA)

 

3 Months + ~10days

Production Bleed 3

 

~200mls

4 Months

Immunisation Boost 4

Freunds Incomplete Adjuvant (IFA)

 

4 Months + ~10days

Production Bleed 4

 

~200mls

5 Months

Immunisation Boost 5

Freunds Incomplete Adjuvant (IFA)

 

5 Months + ~10days

Production Bleed 5

 

~200mls

6 Months

Immunisation Boost 6

Freunds Incomplete Adjuvant (IFA)

 

6 Months + ~10days

Production Bleed 6

 

~200mls

7 Months

Immunisation Boost 7

Freunds Incomplete Adjuvant (IFA)

 

7 Months +~10days

Production Bleed 7

 

~200mls

8 Months

Immunisation Boost 8

Freunds Incomplete Adjuvant (IFA)

 

8 Months + ~10days

Production Bleed 8

 

~200mls

9 Months

Final Immunisation Boost

Freunds Incomplete Adjuvant (IFA)

 

9 Month + ~10days

Final Production Bleed

 

~500mls