A coupling process for improving purity of bacterial minicells by holin/lysin

Drug and gene delivery systems using bacterial minicells have attracted much attention. Here we attempted to enhance the yield and purity of the minicells using a novel method in which autolysin induced in actively metabolizing parent cells led to autolysis. A two-step protocol coupling an autolysin (holin/lysin) step with a conventional centrifugation step achieved a purity of the minicells similar to that resulting from successive six- or seven-step methods.