噬菌体制剂中内毒素脱除方法的比较研究

目的 建立动态浊度法检测噬菌体制剂中内毒素含量, 并探索不同方法对制剂中内毒素的脱除效果。方法 采用动态浊度法检测噬菌体制剂中内毒素含量, 然后采用凝胶层析、超滤及活性炭吸附法对噬菌体制剂进行处理, 检测处理前后噬菌体制剂中内毒素含量及噬菌体效价的变化。结果 基于动态浊度法,建立了内毒素标准曲线回归方程lgT=-0.2732 lgC+3.149, 其线性浓度范围为0.05-10.0 EU/mL,相关系数│r│=0.9986, 满足可靠性分析要求；凝胶层析、超滤法和活性炭吸附法均能有效脱除噬菌体制剂中的内毒素, 脱除率分别为92.6%, 97.1%和99.1%, 但活性炭吸附法选择性差, 噬菌体效价显著降低。结论 动态浊度法能灵敏检测噬菌体制剂中的内毒素含量,凝胶层析和超滤法可作为噬菌体制剂内毒素脱除的可行策略。

Comparative study on endotoxin removal methods from phage preparations

Objective To establish a kinetic turbidimetric assay for detecting endotoxin in bacteriophage preparations and explore the effect of different methods on endotoxin removal. Methods The endotoxin content in the phage preparation was detected by dynamic turbidity method; The phage preparations were processed by gel chromatography, ultrafiltration and activated carbon adsorption. The changes of endotoxin content and phage titer before and after treatment were detected.. Results Based on dynamic turbidity method, the regression equation of endotoxin standard curve was lgT=-0.2732 lgC+3.149 , with a correlation coefficient of 0.9986, and the measurable concentration ranged from 0.05 to 10.0 EU / mL, which met the requirements of reliability analysis. Gel chromatography, ultrafiltration and activated carbon adsorption can effectively remove endotoxin from phage preparations, and the removal rate is 92.6%. 97.1% and 99.1%, but the selectivity of activated carbon adsorption is poor, and the phage titer is significantly reduced. Conclusions Kinetic turbidimetric assay can be used to detect endotoxin in bacteriophage preparations; gel chromatography and ultrafiltration can be used as a feasible strategy for endotoxin removal in phage preparations.