Isolation of Antigenic Fractions from Bordetella pertussis by means of Lysozyme

Abstract

A CONSIDERABLE number of Gram-negative bacteria have been broken up in recent years by means of lysozyme^1–4. The results of lysozyme treatment of Bordetella pertussis and the analysis of the resulting antigenic fractions form the subject of this communication. Suspensions (phase I), obtained from 24-h-old liquid shaken cultures, were used. After washing them with distilled water, we adjusted their germ count to 4 to 6 × 10¹⁰/ml. with reference to the International Standard for Opacity. We determined the conditions under which lysis would least impair antigenicity and found that our purpose would be best served by using a combined treatment with crystalline lysozyme (100 µg/ml.) and versene (200 µg/ml.) in a 0.03 M tris buffer of pH 8.0 under continuous stirring in a water bath of 37° C for 2–3 h.