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; 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, 超滤法可作为噬菌体制剂内毒素脱除的可行策略。
Objective To establish a method for detecting endotoxin in bacteriophage preparations by kinetic turbidimetric assay 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, and 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 the 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. Ultrafiltration and activated carbon adsorption could effectively remove endotoxin from phage preparations, and the removal rate was 97.1% and 99.1%, but the selectivity of activated carbon adsorption was poor, and the phage titer was significantly reduced. Conclusions Kinetic turbidimetric assay can be used to detect endotoxin in bacteriophage preparations, and ultrafiltration can be used as a feasible strategy for endotoxin removal in phage preparations.
标题：噬菌体制剂中内毒素脱除方法的比较研究
英文标题：Comparative study on endotoxin removal methods from phage preparations
作者：
吕可 大连海洋大学食品科学与工程学院
赵前程 大连海洋大学食品科学与工程学院
李萌 大连海洋大学食品科学与工程学院
程少峰 大连海洋大学食品科学与工程学院
姚雪 大连海洋大学食品科学与工程学院
马永生 大连海洋大学食品科学与工程学院
中文关键词:噬菌体制剂,内毒素,凝胶层析法,活性炭吸附法,超滤法,动态浊度法,
英文关键词:phage preparation,endotoxin,gel chromatography,active carbon adsorption,ultrafiltration,dynamic turbidity method,
发表日期：2020-07-27
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