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84 Day Fusion Protein Protocol - in Rabbits

Our experience shows that for certain Fusion Protein Antigens, this protocol works well.  Please note that the route of immunizations include two intradermal injections and one intravenous injection.  Please keep in mind for IV injections, the antigen supply must easily pass through 23 gauge needle and be in PBS buffer.  This protocol is not suitable for antigen on gel.

Day 0 -

Preimmune Bleed, 3 to 4ml of sera. Preimmune bleed is stored until the first immune bleed is sent. 

Inject 200ug intradermal in Complete Freund's Adjuvant (FCA).   Antigen supply should be 0.8mg/ml or higher.

Day 28 -

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

Day 42 -

Inject 50ug subcutaneous with IFA.

Day 56 -

Small Bleed (3 to 4ml sera)

Inject 25ug subcutaneous with IFA.
Inject 25ug intravenous without adjuvant.

Day 63 -

Large Bleed, about 15ml sera.  This bleed will be sent to you along with the preimmune bleed and the day 56 bleed 

Day 70 -

Inject 50ug subcutaneous with IFA

Day 84 - Select one of the three options below:

For CLOSED projects - Large bleed at day 84 - rabbit is then euthanized / discard (lowest cost).  The bleed is then sent to you.

For CLOSED projects - The rabbit is exsanguinated (final bleed) at day 84. The exsanguination sera is then sent to you. 

For OPEN projects - Large Bleed - This bleed is then sent to you.

After Day 84 for OPEN projects -

You may select one of the project extensions below.  If you do not specify a project extension, we will automatically begin Extension A.  Keep in mind that all procedures and maintenance after day 84 will be charged according to the Custom Polyclonal Antibody Production Price Schedule.

Extension A:

We will boost every four weeks with 25-50ug of antigen subcutaneous with IFA. We will do a large 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 sera are needed, we can start production bleeds. We do production bleeds every seven days.  We will continue with production bleeds until you contact us or the health of the rabbit requires us to stop production bleeds.  The volume of serum collected for large or production bleeds depends on the weight and the general health of the Rabbit. We generally take 25-45ml of blood yielding 13 to 20ml of serum.    During this time, we boost the rabbits every four weeks with 25-50ug of antigen subcutaneous 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 84.  Extension C is hold without boosting and bleeding.  We will hold until you tell us to do something else. 



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 rabbit is anesthetized and the terminal bleed is taken.   For best results, rabbits should be boosted 10 to 21 days before exsanguination.
2.      Discard - termination without sera - the rabbit 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.

Custom Extension:

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