百日咳杆菌黏着素的纯化及生物学特性研究

Study on purification and biological characteristics of pertactin from Bordetella pertussis

目的建立从天然百日咳杆菌中提纯百日咳杆菌黏着素（pertactin，PRN）的方法，研究PRN的生物学特性并确定PRN在无细胞百日咳疫苗中的作用。方法百日咳杆菌CS株于发酵罐培养后，经热浸提、硫酸铵分级沉淀及离子交换层析得到纯化PRN，根据《中华人民共和国药典》2005年版三部的要求，对纯化PRN进行纯度检测及生物学特性研究。结果各项检测显示，纯化PRN的纯度在95％以上，其相对分子质量和等电点与标准PRN一致。纯化PRN对小鼠具有保护效力，且其保护效力随PRN免疫浓度的增加而增强。未检出纯化PRN的特异性毒性和异常毒性。结论建立了从天然百日咳杆菌中提纯PRN的方法，而且纯化PRN具有免疫原性和保护效力。

Objective To establish a method for purifying pertactin from Bordetella pertussis, study its biological characteristics and confirm its role in acellular pertussis vaccine. Methods Bordetella pertussis CS strain was cultured in a fermenter, and purified pertactin was obtained by hot lixiviation, ammonium sulfate fractional precipitation and ion-exchange chromatography. Purity and biological characteristics of the purified pertactin were studied according to requirements of Pharmacopoeia of the People＇s Republic of China 2005 Volume lII. Results The purity of the purified pertactin was above 95%, relative molecular mass and isoelectric point of the purified pertactin were identical with those of standard pertactin. Protective potency of the purified pertactin was observed in mice and enhanced with increase of its concentrations. No specific and abnormal toxicities were detected. Conclusions The method for purifying pertactin from Bordetella pertussis is established, and purified pertactin has immunogenicity and protective potency.