Polyclonal Antibody Production

Polyclonal Antibody Production

Approved June 25, 1998    (Download) 

Background: Reliance has traditionally been placed on the use of Fruend’s Complete Adjuvant (FCA) as an adjuvant, or antibody production enhancer, for producing polyclonal antibodies in animals. FCA has been used for many years in several species, mice and rabbits being the most frequent. Inflammation results from the use of FCA and reactions vary according to the anatomical site in which it is placed, the volume of antigen used, and the purity of the antigen. Severe reactions to FCA sometimes occur following inoculations in mouse foot pads and in rabbits, although such reactions can be minimized when very small volumes are given. There are reports that small subcutaneous injections of FCA in the lower leg are equal in effectiveness to inoculations in the foot pad and cause much less reaction. Discontinuance of the use of FCA has been advocated; however, other adjuvants are generally not as effective as FCA. FCA remains widely preferred as the best adjuvant for use in stimulating antibody production.

Administering FCA and antigen aseptically and in smaller doses at multiple rather than a single sites over a wide body area to stimulate a greater spectrum of the immune system has resulted in acceptable levels of antibody production without the development of painful lesions.


Policy: Use of FCA in polyclonal antibody production is permitted if the investigator provides scientific justification for its use, rather than less irritating adjuvants. FCA , if used for antibody production, should be divided into multiple small doses given at no more than 10 widely dispersed sites covering the four quadrants of the body. Each injection site should receive no more than 0.1 ml and subcutaneous injection is preferred. Precautions that must be observed include aseptic preparation of the injection site and sterility of the inocula. FCA should be used in only the first injection; subsequent injections requiring an adjuvant must use Freund’s incomplete or another appropriate adjuvant. Use of the foot pad as an injection site is generally unacceptable. Waivers to perform foot pad injections may be granted by the ACC if investigators are able to demonstrate that the use of preferred alternatives is unsatisfactory and there is compelling scientific justification to do so. In cases where such a waiver may be granted, special soft bedding must be provided to reduce the pain of lesions that may occur on the feet.

Documentation must be provided by the investigator that competency of project personnel to inject FCA has been demonstrated to the satisfaction of the attending veterinarian or by a qualified individual designated by the attending veterinarian.