Abstract
Three types of complement-fixing antigen prepared from yolk sac cultures of the agent of lymphogranuloma venereum have been described; viz. (1) resuspended high speed sediment, 1 (2) soluble antigen in high speed supernates, 2 (3) whole suspensions inactivated with urea or ether. 3 The third type was approximately four times as active as high speed sediment.
Attempts by one of the authors (Nigg) to find a satisfactory preservative for lymphogranuloma venereum complement-fixing antigens, showed that phenol enhanced the activity of the antigen specifically since such phenolized antigens did not react with normal sera. This observation was studied further and is the subject of this note.
Table I shows the complement-fixing activity of a 10% yolk sac suspension treated with (a) 2% urea, (b) 0.25% phenol and (c) 0.5% phenol. Preparations were stored at icebox temperatures. Two separate fractions of suspension 56 when treated with 0.25% phenol failed to show increased activity when tested 8 to 30 days later. Two fractions treated with 0.5% phenol were 4 and 8 times respectively as active as the urea-treated antigen.
While 0.25% phenol had little enhancing effect at icebox temperatures, Table II shows approximately a 4-fold increase in activity when these suspensions were subjected to the following temperatures: (a) 37°C for 6 weeks (not tested earlier), (b) 56°C for 48 hours, and (c) boiling water for 10 minutes. Furthermore, the centrifuged (2,000 rpm for 10 minutes) supernates of the heated suspensions were almost as active as the whole heated suspensions. This represents a considerable purification since the slightly opalescent supernates, after removal of tissue debris and coagulated protein, were almost as active as the whole heated turbid suspensions.
That the phenol enhancement is apparently specific is shown in diagnostic tests (Table III) with positive and negative sera, comparing 2 phenolized antigens with urea-treated antigen.
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