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口蹄疫灭活疫苗蛋白质含量测定操作程序的确立
引用本文:马丽,赵启祖.口蹄疫灭活疫苗蛋白质含量测定操作程序的确立[J].中国兽药杂志,2014,48(5):11-16.
作者姓名:马丽  赵启祖
作者单位:中国兽医药品监察所,中国兽医药品监察所
基金项目:公益性行业(农业)科研专项(201303046-07)
摘    要:为优化用于口蹄疫灭活疫苗蛋白质含量测定的改良Lowry法,进而确立口蹄疫灭活疫苗蛋白质含量测定的操作程序,探索了有机溶剂破乳剂、酚红以及丙酮沉淀对测定结果影响。结果表明:样品中含酚红和有机溶剂均导致测定值较标准值高;有机溶剂破乳后,水相样经过丙酮沉淀测定值较标准值低;丙酮直接沉淀疫苗后测定蛋白质值与标准值符合度最高,丙酮沉淀回收率随蛋白浓度升高而升高,回收率在90%~100%之间。试验首次确立了改良Lowry法检测口蹄疫灭活疫苗中蛋白质含量的操作程序为丙酮直接沉淀疫苗后测定蛋白质浓度。并成功应用于口蹄疫灭活疫苗蛋白质含量的测定。

关 键 词:改良Lowry法  口蹄疫灭活疫苗  蛋白质含量
收稿时间:2014/3/24 0:00:00
修稿时间:2014/4/24 0:00:00

Establishment of the operating procedures about Protein Quantitation Assay of FMD Inactivated Vaccine
MA Li,ZOU Xing-qi,ZHU Yuan-yuan,ZHAO Qi-zu.Establishment of the operating procedures about Protein Quantitation Assay of FMD Inactivated Vaccine[J].Chinese Journal of Veterinary Drug,2014,48(5):11-16.
Authors:MA Li  ZOU Xing-qi  ZHU Yuan-yuan  ZHAO Qi-zu
Institution:China Institute of Veterinary Drug Control,
Abstract:In order to optimize the modified Lowry method and then set up operating procedures for quantify protein of FMD inactivated vaccines, the influence of organic solvent demulsified, phenol red and acetone precipitating on the measurement results was explored. The measurement results indicated that phenol red or organic solvents in samples would lead to measurement results higher than standard values. The measurement results of samples which were demulsified by organic solvent and then the aqueous phase were precipitated by acetone was lower than standard values. The measurement results of samples which were precipitated directly by acetone were most close to standard values. The recovery of acetone precipitation increased with elevated concentration of protein, which was between 90% and 100%.The operating procedures of modified Lowry protein assay which was used to quantify protein of FMD inactivated vaccine were established for the first time. The procedures were that the vaccines were dealed with actone first, and then the protein that precipitated by actone was suspended and quantified. The assay was successfully applied in Protein quantitation of FMD inactivated vaccines.
Keywords:modified Lowry protein assay  FMD inactivated vaccine  protein quantitation
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