Rationale:

Production of polyclonal antibodies at UT Southwestern will be consistent with standards developed by this institution to meet the goals of its Animal Care and Use Program.

Definitions:

Polyclonal antibodies are produced when a large number of B lymphocytes produce antibodies against different parts of the same antigen or multiple antigens.  Typically, animals are immunized multiple times to elicit a strong antibody response, then bled so that immune serum can be collected and used in experiments.  The USDA in Animal Care Policy # 11 includes the use of Complete Freunds Adjuvant (CFA) as an example of a procedure that can be expected to cause more than momentary or slight pain.

Policy:

• Polyclonal antibody production methods must be described in the PIs protocol.
• If CFA will be used in a protocol, the PI must:
  1.  Conduct a literature search for alternatives to the use of CFA in the animal protocol.  (Examples of alternative adjuvants include: Titermax, Ribi, and Incomplete Freunds Adjuvant.);
  2.  Scientifically justify the use of CFA in the animal protocol; and
  3. Provide eye protection for employees using CFA.
•  CFA must only be used once with booster immunizations employing Incomplete Freund’s Adjuvant or other appropriate adjuvant. 
• The concentration of mycobacteria in CFA must be 0.5 mg/ml or less.

Applicability:

This policy applies to polyclonal antibody production performed at UT Southwestern.

Procedures:

The published ILAR journal (Volume 46, no. 3, 2005) is an excellent reference for polyclonal antibody production.  This journal can be accessed using the following web address:

http://dels.nas.edu/ilar_n/ilarjournal/46_3/html/

•  Subcutaneous injection routes are acceptable methods for antigen delivery in rodents.  Subcutaneous and intradermal routes are acceptable methods for antigen delivery in rabbits and larger animals.  Other routes must be scientifically justified in the protocol.
• Injections must be limited to 4 sites or less for both initial and booster immunizations.
• Prior to subcutaneous immunization, the skin must be wiped clean with 70 % isopropyl alcohol or an appropriate disinfectant such as chlorhexidine solution.
• Prior to intradermal immunization, the hair must be clipped from the injection sites and prepared with a surgical scrub such as betadine or chlorhexidine.
• Immunization injections should be spaced 3-6 weeks apart using small volumes.
• Immunizations in non-rodents should be spaced over the four quadrants of the animal (behind the shoulder and in front of the hind leg, left and right side).  Bilateral injections at the base of the tail in rodents effectively stimulates an immune response.
• Injection sites should be routinely monitored for the development of pathology such as granuloma formation or skin ulceration.
• The veterinary staff shall be notified and consulted regarding treatment of any lesions that develop.  Lesions identified by the veterinary staff will be treated as appropriate after consult with the investigative group.
• Periodic survival blood collections must not exceed 1% of the animals body weight in a two week period.

Exceptions:

Exceptions to this policy may be granted by the IACUC if scientifically justified in an approved protocol.

Contact Information:

For any additional questions, please contact the IACUC Manager at 5-6420.