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Guide for Investigators: Immunizing rabbits
Standard Operating Procedure, Veterinary Clinical Services
Section of Comparative Medicine, Yale University

Purpose:

To immunize rabbits for polyclonal antibody production.

Materials:
Bullet. 3.0 cc syringe
Bullet. 20 gauge needle
Bullet. gauze
Bullet. hydrogen peroxide

Procedure:

  1. Remove rabbit from cage and place on table. Proper technique in grabbing and carrying a rabbit is important to prevent vertebral fractures. Grab the rabbit's skin in the shoulder region and place its head underneath your arm. Support the rabbit's legs with the hand of the arm that the head is tucked under. Alternatively, grab the rabbit's skin with both hands (over both the shoulder and the rump region).
  2. Mix antigen with adjuvant as recommended by manufacturer. Freund's adjuvant (CFA) commonly causes inflammatory reactions, such as, granulomas and ulceration. Alternative adjuvants should be used whenever possible.
  3. The rabbit's back must shaved for primary immunization. Record primary injection sites in the animal's record. If the rabbit is very nervous it may be necessary to sedate it with Acepromazine 2mg/kg (see Blood Collection SOP for instructions on how to administer).
  4. Each subcutaneous or intradermal injection site must not exceed 0.1cc and no more than a total of 10 sites may be injected.
  5. The injections must be confined to an area on the rabbits back that is:
    Bullet. No higher than the shoulder blades.
    Bullet. No lower than the pelvic bone.
    Bullet. Laterally no lower than midway down torso
    Bullet. Intraperitoneal and intramuscular injections and those on the extremities are not allowed.
  6. If injecting an antigen contained in gel, document each boost on the rabbit outline in the record.
  7. It is recommended that rabbits be immunized 2-4 times every 21 days, depending on the anticipated immunogenicity of the antigen/adjuvant combination. Alterations to this schedule and anticipated end-points for the experiment need to be addressed in the IACUC protocol.
  8. Clean up any blood on the fur with hydrogen peroxide and gauze.

Veterinary approval Date: 11/21/2003
Endorsed by the IACUC Date: 12/10/2003

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