Bandiera Silvio, Lebas Annabelle, Canizares-Martinello Livia, Guinchard Fabien, Lyonnais Corinne, Perrin Sophie, Nicolas Marine, Uhlrich Sylvie, Chabaud-Riou Martine
Sanofi Pasteur, Campus Mérieux, 1541 Avenue Marcel Mérieux, 69280, Marcy l'Etoile, France.
Biologicals. 2019 Sep;61:15-21. doi: 10.1016/j.biologicals.2019.08.002. Epub 2019 Aug 23.
The diphtheria toxoid (DT), tetanus toxoid (TT), and acellular pertussis (aP) single immunogenicity assay (DTaP SIA) is a Luminex®-xMAP®-bead-based multiplex immunoassay for estimating the potency of DTaP pediatric combination vaccines in guinea pigs. This manuscript describes the validation of this assay for the simultaneous quantitation of anti-diphtheria toxoid (anti-DT), anti-tetanus toxoid (anti-TT), anti-pertussis toxoid (anti-PTxd), and anti-filamentous hemagglutinin (anti-FHA) antibodies in guinea pig serum following injection of a DTaP vaccine formulation. The results were expressed in arbitrary units/mL (AU/mL) using reference serum for comparison. Specificity was demonstrated by ≥ 75% homologous and ≤25% heterologous inhibition for all the antigens. The results were linear for anti-DT, anti-TT, anti-PTxd and anti-FHA antibodies. Accuracy was demonstrated with recovery of between 80% and 120% for all four antibodies. The relative standard deviation of repeatability was ≤20%. The results demonstrate that this SIA can be used for the linear, accurate, and precise simultaneous detection of all four antibodies, based on both the ICH Q2 and the EMA guidelines on bioanalytical method validation.
白喉类毒素（DT）、破伤风类毒素（TT）和无细胞百日咳（aP）单一免疫原性测定（DTaP SIA）是一种基于Luminex®-xMAP®微珠的多重免疫测定法，用于评估豚鼠中DTaP儿科联合疫苗的效力。本手稿描述了该测定法的验证情况，该测定法用于在注射DTaP疫苗制剂后同时定量豚鼠血清中的抗白喉类毒素（抗-DT）、抗破伤风类毒素（抗-TT）、抗百日咳毒素（抗-PTxd）和抗丝状血凝素（抗-FHA）抗体。使用参考血清进行比较，结果以任意单位/毫升（AU/mL）表示。所有抗原的同源抑制率≥75%且异源抑制率≤25%，证明了该测定法的特异性。抗-DT、抗-TT、抗-PTxd和抗-FHA抗体的结果呈线性。所有四种抗体的回收率在80%至120%之间，证明了该测定法的准确性。重复性的相对标准偏差≤20%。结果表明，根据国际人用药品注册技术协调会（ICH）Q2和欧洲药品管理局（EMA）关于生物分析方法验证的指南，该SIA可用于对所有四种抗体进行线性、准确和精确的同时检测。
