Aggregation and Activity of Forssman Hapten in Dilute Aqueous Solutions

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Abstract
Summary: The results obtained in this study support the hypothesis that activators in the inhibition assay act as dispersing agents for the Forssman hapten. When phrenosin was used in this role, the activation was not persistent and activity decreased significantly when the aqueous system was allowed to stand for 24 hr. The possibility that the loss of activity was due to atmospheric oxidation of the hapten was eliminated by means of storage experiments under conditions avoiding such oxidation. Centrifugation of solutions that had reached a low activity on standing lowered the activity even more, indicating that the dispersed material had coalesced to form larger particles. The aggregates were shown to be composed of both phrenosin and hapten. Activity was restored to solutions whose activity had thus declined by lyophilization, re-solution in methanol, and dilution in the aqueous assay system. This treatment apparently results in a dispersion state similar to that in the original solution. Phrenosin served as an activator for the hapten by delaying its micellar formation. Several other materials were used as activators but only one, a fraction from the purification procedure, was as effective as phrenosin.

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