Materials 21 Immunizations

1. Freund's complete and incomplete adjuvants: FCA is a suspension of heat-killed Mycobacterium tuberculosis in oil. This should be shaken well before use. FIA is the oil without the bacteria. Freund's adjuvants should be mixed with the aqueous immunogen solution/ suspension at a ratio between 1:2 and 2:1 and mixed until an emulsion is formed. This is most easily achieved by passage back and forth between two glass syringes connected by a three-way luer fitting. After a short time (1-5 min) the mixture should become noticeably "stiffer" and may then be administered.

2. Potassium aluminium sulphate (Alum) AlK(SO4)2T2 H2O precipitates: the aqueous immunogen solution/suspension is mixed with 0.3 vol 10% alum. The pH is then adjusted to about 8.0 with sodium hydroxide solution and the resultant precipitate washed in 0.154 M NaCl solution and administered.

3. Acetone precipitates: the aqueous immunogen is precipitated with 4.5 vol acetone at -20°C. The precipitate is collected by centrifuga-tion at 10,000g at room temperature, washed in 80% acetone and air-dried. The pellet is resuspended in saline using a Dounce™ homogenizer and then administered directly or in association with alum or Freund's adjuvants.

6. Disposable three-way luer fitting taps.

7. Peptide-conjugate/purified protein immunogen.

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