Establishment of A Laboratory Method for Removing Endotoxin from Phage Preparations
Endotoxins are outer membrane lipopolysaccharides released when Gram-negative bacteria are lysed by phages.Endotoxin is the main contaminant in bacteriophage preparation,which may cause severe immune reaction.Therefore,endotoxin removal is very important for phage applications.In this paper,we used dialysis,ultrafiltration and a combination of the two to process the phage preparations.The concentration of endotoxin and bacteriophage titer were detected before and after different processes.The results show that all three methods can effectively remove endotoxins in phage preparations.The dialysis method showed the best effect of removing endotoxin in 48 hours,and the removal rate was over 99.9％.In addition,the phage recovery rates obtained by ultrafiltration method was significantly lower than that obtained by dialysis method,and the endotoxin content per plaque forming unit(PFU)is higher.The combination of ultrafiltration and dialysis saves time at the expense of partial recovery,and the removal rate of endotoxin can reach more than 99％.In summary,this work established a method for removing endotoxins from phage preparations through dialysis.Compared with traditional ultrafiltration,dialysis has the advantages of simple operation,low cost,and high recovery rate.It can be used as a feasible method for removing endotoxins from phage preparations in the laboratory.
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维普
