Medicines and Healthcare products Regulatory Agency, South Mimms, UK.
Medicines and Healthcare products Regulatory Agency, London, UK.
ALTEX. 2024;41(4):588-604. doi: 10.14573/altex.2401171. Epub 2024 Aug 23.
Antigen identity, quantity and integrity are key factors to be evaluated as part of consistency testing of tetanus vaccines. Here we have developed a monoclonal antibody sandwich ELISA to measure the relative amount and quality of tetanus toxoid (TTxd) in human and animal tetanus vaccines. The ELISA is highly specific, has good dilutional linearity, and is suitable for detecting TTxd in a range of different products. We have demonstrated the ability of the assay to discriminate between batches of different content, using vaccine batches that had been prepared to contain differing amounts of TTxd, and of different quality, using samples of non-adjuvanted TTxd that had been exposed to sonication and final lot vaccines that had been exposed to heat or oxidative stress. We have also demonstrated successful transfer of the method to other laboratories and have shown that different tetanus antigen materials may be able to serve as a reference antigen for standardization of the method. The results show this test has the potential to play a key role in a control strategy no longer including an in vivo potency test.
抗原的种类、数量和完整性是破伤风疫苗一致性检测的关键评估因素。本研究开发了一种用于测量人用和动物用破伤风疫苗中破伤风类毒素(TTxd)相对含量和质量的单克隆抗体夹心 ELISA 法。该 ELISA 法具有高度特异性、良好的稀释线性,适合检测不同产品中的 TTxd。我们使用制备时 TTxd 含量不同和质量不同的疫苗批次,证明了该方法有能力区分不同含量和质量的批次,也使用了经过超声处理的非佐剂 TTxd 样品和经过热或氧化应激处理的最终批疫苗来证明。我们还成功地将该方法转移到其他实验室,并表明不同的破伤风抗原材料可能能够作为该方法标准化的参考抗原。结果表明,该检测方法有可能在不再包括体内效力测试的控制策略中发挥关键作用。