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Home  >  PRF&L Traditional Protocols  >  Chicken custom antibody production

TRADITIONAL 70 DAY POLYCLONAL ANTIBODY PRODUCTION - in Chickens


Day 0 -

Preimmune Bleed, generally 5 to 10ml plasma. Preimmune bleed is stored until the first immune bleed is sent.

Preimmune Egg, one egg is saved prior to immunizations.  Pre egg is cracked and yolk stored at -80C.  Additional pre eggs can be saved at additional cost if requested.

Inject 200ug intramuscular in Complete Freund's Adjuvant (CFA).   We will inject intramuscular and subcutaneous if the concentration of antigen is less than 0.4mg/ml. Acrylamide gel can only be injected subcutaneous.

Day 14 -

Inject 100ug intramuscular in Incomplete Freund's Adjuvant (IFA)

Day 28 -

Inject 100ug intramuscular with IFA.

Day 42 -

Bleed (5 to 10ml plasma) We will ship via Federal Express the preimmune bleed and this first immune bleed.  It is then up to the investigator to test the serum, determine the quality of antibodies, and let us know if any changes are necessary in the protocol.

Day 56 -

Inject 50ug intramuscular with IFA

Day 70 -

Bleed (5-10ml sera). We will ship via Federal Express this small bleed. It is up to the investigator to test the serum, determine the quality of antibodies, and let us know if any changes are necessary in the protocol.

After Day 70 -

You may select one of the project extensions below.  If you do not specify a project extension, we will automatically begin Extension A.

Extension A:

We will boost every four weeks with 25-50ug of antigen subcutaneous or intramuscular with IFA. We will do a bleed two weeks after each boost and ship. We will continue to boost and bleed every four weeks until you contact us. 

Extension B:

When the antisera contains sufficient antibody titer and large volumes of plasma are needed, we can start production bleeds. We do production bleeds every seven days.  We will continue with the production bleeds until you contact us or the health of the chicken requires us to stop production bleeds.   The volume of plasma collected for production bleeds depends on the weight and the general health of the chicken.  We generally take 10-25ml of blood yielding 5 to 12ml of plasma.   During this time, we boost the chickens every four weeks with 25-50ug of antigen subcutaneous or intramuscular with IFA.   You may also request that we not do any boosts after the initial immunization scheme outlined above since some antigens and some animals may respond negatively to boosts during the period of peak antibody activity. Boosts may be required only when titer drops. It is up to the investigator to decide and let us know if we should discontinue boosts as our standard schedule provides for boosts every 28 days.    Shipping for production bleeds is every four weeks unless you request otherwise.

Extension C:

Some investigators prefer to "wait and see." after day 70.  Extension C is hold without boosting and bleeding.  We will hold until you tell us to do something else.  Our experience shows that the peak antibody titer occurs at an average of 120 days, however we have experienced the onset of peak activity as early as six weeks and as long as a year. If results are not as you expect, please consult with us and we can make recommendations on how to change your protocol.

Termination:

When you have collected enough antisera, you will need to contact us to terminate the project. You may request that we terminate by:
1.      Exsanguination - termination with sera - the chicken is anesthetized and the terminal bleed is taken.  For best results, chickens should be boosted 10 to 21 days before exsanguination.
2.      Discard - termination without sera - the chicken is humanely euthanized.
        
When requesting termination, you must confirm the species and animal number. Written confirmation by email or fax may be requested for terminal procedures.

Extension E -  E is for Egg

Large amounts of antibody can be harvested from chicken egg yolks.  Upon request, we can collect as many eggs as you need.  You need to contact us to request immune egg collection.  You may request that we collect a target number of eggs or simply collect all the eggs the chickens lay.  We offer a egg purification service.  If you prefer to process your own yolks, we will crack the eggs and send the yolks.
Many investigators prefer to test bleeds and then use the eggs for purification.

Custom Extension:

You may design your own project extension.  Let us know what we can do for you!