Cheung Chung Y, Dubey Sitara, Hadrovic Martina, Ball Christina R, Ramage Walter, McDonald Jacqueline U, Harvey Ruth, Hufton Simon E, Engelhardt Othmar G
Vaccines Division, Scientific Research & Innovation, Medicines and Healthcare Products Regulatory Agency, South Mimms, Potters Bar EN6 3QG, UK.
Biotherapeutics Division, Scientific Research & Innovation, Medicines and Healthcare Products Regulatory Agency, South Mimms, Potters Bar EN6 3QG, UK.
Vaccines (Basel). 2022 Sep 5;10(9):1473. doi: 10.3390/vaccines10091473.
Inactivated vaccines are the main influenza vaccines used today; these are usually presented as split (detergent-disrupted) or subunit vaccines, while whole-virus-inactivated influenza vaccines are rare. The single radial immune diffusion (SRD) assay has been used as the gold standard potency assay for inactivated influenza vaccines for decades; however, more recently, various alternative potency assays have been proposed. A new potency test should be able to measure the amount of functional antigen in the vaccine, which in the case of influenza vaccines is the haemagglutinin (HA) protein. Potency tests should also be able to detect the loss of potency caused by changes to the structural and functional integrity of HA. To detect such changes, most alternative potency tests proposed to date use antibodies that react with native HA. Due to the frequent changes in influenza vaccine composition, antibodies may need to be updated in line with changes in vaccine viruses. We have developed two ELISA-based potency assays for group 1 influenza A viruses using cross-reactive nanobodies. The nanobodies detect influenza viruses of subtype H1N1 spanning more than three decades, as well as H5N1 viruses, in ELISA. We found that the new ELISA potency assays are sensitive to the nature of the reference antigen (standard) used to quantify vaccine antigens; using standards matched in their presentation to the vaccine type improved correspondence between the ELISA and SRD assays.
灭活疫苗是目前使用的主要流感疫苗;这些疫苗通常以裂解(去污剂破坏)或亚单位疫苗形式呈现,而全病毒灭活流感疫苗则较为罕见。几十年来,单向辐射免疫扩散(SRD)试验一直被用作灭活流感疫苗效力检测的金标准;然而,最近人们提出了各种替代效力检测方法。一种新的效力检测方法应该能够测量疫苗中功能性抗原的量,对于流感疫苗来说,就是血凝素(HA)蛋白。效力检测还应该能够检测由于HA的结构和功能完整性改变而导致的效力损失。为了检测这种变化,迄今为止提出的大多数替代效力检测方法都使用与天然HA反应的抗体。由于流感疫苗成分经常变化,抗体可能需要根据疫苗病毒的变化进行更新。我们使用交叉反应纳米抗体开发了两种基于ELISA的甲型流感病毒1组效力检测方法。这些纳米抗体在ELISA中能够检测跨越三十多年的H1N1亚型流感病毒以及H5N1病毒。我们发现,新的ELISA效力检测方法对用于定量疫苗抗原的参考抗原(标准品)的性质敏感;使用与疫苗类型呈现方式匹配的标准品可提高ELISA和SRD检测之间的一致性。