STUDIES ON IN VITRO ANTIBODY PRODUCTION: I. THE ESTABLISHMENT OF THE METHOD

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Abstract
For the study of in vitro antibody production, methods have to be established which are sensitive and specific enough to demonstrate minute amounts of antibodies.Spleen cell suspensions from hyperimmunized rabbits (secondary phase) were incubated in completely synthetic medium. The amount of antibody in the culture fluid supernatants was assayed by the following methods: Neutralization of C. perfringens toxin using a modified lecitho-vitellin test. The test could demonstrate as little as 0.0015 antitoxic units (AU)/ml. In tissue culture systems antitoxin titers up to 0.1 AU/ml were observed. Neutralization of the dermonecrotic and lethal action of diphtheria toxin. This method detected 0.001 AU/ml. In experiments with tissue cultures antitoxin titers of 0.25 AU/ml could be demonstrated.Antibodies labelled with C14glycine were used in other experiments. To demonstrate labelled antibodies the coprecipitation method was used.Our experiments showed that antibodies produced in vitro can(a) inhibit the lecithinase activity of C. perfringens toxin,(b) neutralize the dermonecrotic and lethal action of diphtheria toxin, and(c) participate in precipitation.

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