Guidelines for Producing Monoclonal Antibodies (MAb) in Mice

I. REFERENCES

Based on these references the following recommendations and requirements are made for the production of MAb in mice.

II. RECOMMENDATIONS

  1. Studies have shown that an F1 hybrid (Swiss, female X BALBIc, male) produced 3-4 times the ascites fluid compared to the BALB/c parents. Investigators should consider using such hybrids as a means of reducing the number of animals required to produce the desired quantity of antibody.
  2. Since the production of ascites fluid is better in males than females, male mice are the preferred sex for MAb production.
  3. Mice should be 43-78 days old.

III. REQUIREMENTS

  1. Pertinent dates must be recorded on the cage card as required by the "Cuide41. The following dates shall be recorded: 1). injection of pristane, 2). injection of hybridoma cells, 3), initial tap, and 4). date of final tap and euthanasia. Cage cards must be retained as part of the research records for subsequent review by the Animal Care and Facilities Committee, and any accrediting boards.
  2. Each mouse should be treated with no more than .5 ml of pristane intraperitoneally.
  3. The priming period for pristane should be 14 days prior to the injection of hybridoma cells.
  4. Generally, collection periods will be on the order of 6 days from injection of hybridoma cells until final collection and euthanasia. Considerable variation may be experienced and mice showing no signs of distress may be held for longer periods. Conversely, mice may become sick and require early euthanasia.
  5. The number of taps shall be limited to two, with the final tap being conducted under anesthetic and followed by euthanasia. Observations have shown an increase in signs indicating stress and adverse effects after the second tap as well as a decline in the volume of asites fluid collected.
  6. Mice shall be observed at least twice daily (at the beginning and end of each day, Including weekends) for signs of distress. Signs such as excessive abdominal enlargement, rough hair coat, difficulty breathing, and a lack of mobility shall be cause for making the final tap followed by euthanasia. It should be remembered that some mice may exhibit signs of distress and require euthanasia even before they are ready for an initial tap. Such animals should be euthanitized at once.

IV. EXCEPTIONS

Investigators wishing to use alternative procedures must provide written justification and receive written approval from the ACFC prior to beginning procedures.