摘要
目的:建立定量检测破伤风类毒素(TT)抗原的双抗体夹心ELISA法,并探索其初步应用。方法:以TT抗毒素为包被抗体,大鼠抗TT多抗为检测抗体,采用相应酶标抗体检测TT抗原含量,建立双抗体夹心ELISA法,并进行方法学验证。结果:TT含量在0~0.062 5 Lf/ml范围内线性关系良好(r>0.99)。该方法与白喉类毒素、百日咳类毒素、百日咳丝状血凝素及百日咳黏附素无明显交叉反应,重复性好,特异性较强,精密度及准确度验证均符合常规质控要求。该法准确检测范围为0.000 625~0.040 000 Lf/ml,定量限度为0.000 625 Lf/ml。采用该方法对TT抗原进行吸附率检测,分别采用该法和絮状单位测定法测定6批TT的絮状含量,2种检测方法高度相关(r=0.94)。结论:建立的定量检测TT的双抗体夹心ELISA法为破伤风疫苗生产过程中TT质量控制提供了有效技术手段。
Objective:To develop a double antibody sandwich ELISA in quantitative determination of tetanus toxoid(TT)antitoxin and to explore its preliminary application.Methods:With TT as enveloping antibody and rat anti-TT multi-antibody as detection antibody,enzyme-labeled antibody was used to detect content of TT antigen,and a double-antibody sandwich ELISA method was established,whose methodology was verified.Results:Best linearity of dose-response curve was found in a range of 0~0.062 5 Lf/ml(r>0.99).The tested result was no apparent cross reactions with Diphtheria toxoid,Pertussis toxoid,Filamantous hemagglutinin and Pertactin,respectively,which showed an excellent repeatability and specificity in reaction with TT,in accordance with requirements for conventional quality control measures.Detection range and quantitation limit were determined as 0.000 625~0.040 000 Lf/ml and0.000 625 Lf/ml,respectively.Adsorption rate of TT antigen was detected by this method,and flocculent content in 6 batches of TT toxoid was determined using this method and flocculent unit method,showing a high correlation(r=0.94).Conclusion:The double antibody sandwich ELISA was developed in quantitative determination of TT,which provided an effective technical means in quality control of TT in production process of TT vaccine.
作者
祝传顺
周振发
洪禹
薛莹
姚雷(指导)
朱卫华
ZHU Chuanshun;ZHOU Zhenfa;HONG Yu;XUE Ying;YAO Lei;ZHU Weihua(Beijing Zhifei Lvzhu Biopharmaceutical Co.Ltd.,Beijing Bacterial Vaccine Engineering Research Centre,Beijing 100176,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第10期1228-1232,共5页
Chinese Journal of Immunology
