Polyclonal Antibody Development PDF Print E-mail
We offer polyclonal antibody production in rabbits. We recommend at least 100 to 200 micrograms dosage of immunogen for the first immunization. For subsequent immunizations, we then continue to immunize using half the initial dosage. The total amount of immunogen needed is approximately 4 - 5 mg for two rabbits.

Besides purified proteins, we use nitrocellulose or gel containing appropriate amounts of immunogen, conjugated peptides and recombinant proteins. If the size of the antigen is less than 20 kD, we recommend conjugation to a carrier protein, such as KLH.

We collect the first test bleed from the animals 10 days after the third immunization, screen the serum by ELISA to evaluate the titer and send the test bleeds to you for confirmation at your request. Collection of production bleeds begins when the screening data indicates that the animal has produced a high enough titer for your application.

Our Standard Protocol includes:

Day 0: Preimmunization bleed and primary immunization with 100 to 200 micrograms antigen in Freund’s Complete Adjuvant.

Day 21: Second immunization with 50 - 100 micrograms in Freund’s Incomplete Adjuvant

Day 42: Third Immunization with 50 - 100 micrograms in Freund’s Incomplete Adjuvant

Day 52: Test bleed collection & screen by ELISA

After you receive the serum from the last collection on day 52, you have a week to evaluate the antibody titer before deciding to extend the protocol on monthly basis, to place the project on hold, or to terminate the protocol. Thus, there are three options: (1) the project can be terminated with exsanguination (65 -75 ml serum from one rabbit is obtained), (2) the project can be continued on monthly basis or (3) the animals may be on hold on daily basis until a decision is made.

Depending on the titer of the antibody, the client may want Production Serum (about 20 - 25 ml/rabbit). Production serum may be collected preferably in between 10 to 14 days after a boost. Rabbits can be safely bled 2 times per month (yield of serum about 40 ml) until required quantity of serum is achieved. Or, the project can be terminated with exsanguination and about 65 - 75 ml serum can be obtained.

The animals can also be on hold by the customers until a decision is made. If we do not hear from you by the 15th day after shipping serum, we will euthanize the animals.