口蹄疫灭活疫苗蛋白质含量测定操作程序的确立

Establishment of the operating procedures about Protein Quantitation Assay of FMD Inactivated Vaccine

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。

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