Proceedings of the Production of Monoclonal Antibodies Workshop

August 29, 1999 | Bologna, Italy

This document has been adapted with permission from a publication created by the Alternatives Research & Development Foundation (ARDF)

Checklist

IACUC Checklist: Monoclonal Antibody Production (In Vitro vs. In Vivo (Ascites))

  1. Is the production of Monoclonal Antibodies (MAb) scientifically justified? What are the goals of the research/ project that require antibodies?
  2. What quantity of antibody is needed; and over what period of time?
  3. Have the hybridomas to be used been Mouse Antibody Production (MAP) tested whether in vivo or in vitro production will be used? This is critical to protect animal colonies during production and later use of MAb products.
  4. Has there been an adequate attempt to expand the hybridoma in vitro? In vitro MAb production attempts should be required with in vivo (ascites) production only as a justified alternative. See in vitro questions below.

In Vitro

  1. Which in vitro systems have been or will be used? Are these appropriate for the quantities needed?
  2. How was the clone (or clones) selected? Production and activity should be factors in clone selection to increase success of in vitro production.
  3. Have the hybridoma cells been adapted for growth in the preferred media? Can serum free or protein free media be used?
  4. Has use of a core facility or commercial source been considered?
  5. Is the cost reasonable? Although not a primary determinant, is the cost for in vitro production much greater than the cost of in vivo production? What factors were included in cost estimates?
  6. Have all attempts at in vitro production been carried out without an acceptable product? Only if yes, proceed to in vivo questions.

In Vivo (Ascites)

  1. Might in vivo pilot studies be appropriate?
  2. Are the mouse strain and number of animals appropriate?
  3. Is the priming appropriate in terms of type of chemical, amount, and length of time prior to inoculation of hybridoma?
  4. Are guidelines for number and timing of taps well described, and adequate to minimize animal discomfort? In general there should be a three-tap maximum, e.g. two survival taps and one following euthanasia. Also consider a one-tap limit to minimize the occurrence of unrelieved discomfort.
  5. Will animals be monitored adequately, and are clinical signs which constitute criteria for euthanasia well defined?