Moon Sung-Sil, Wang Houping, Brown Kimberly, Wang Yuhuan, Bessy Theresa, Greenberg Harry B, Jiang Baoming
Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Department of Medicine and Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.
J Virol Methods. 2025 Feb;332:115079. doi: 10.1016/j.jviromet.2024.115079. Epub 2024 Nov 26.
To determine the potency of the inactivated rotavirus vaccine (IRV), we developed an enzyme immunoassay (EIA) using a biotin-conjugated RV VP7-specific monoclonal antibody. RV VP7, a pivotal structural protein in the outer capsid layer, governs RV G genotypes and prompts host immune responses, including neutralizing antibodies. This EIA showed high specificity, good linearity, high precision, and high accuracy, with a low limit of detection (LOD) and a limit of quantitation (LOQ) of 0.037 µg/ml RV antigen. The EIA was evaluated and proved suitable for establishing the long-term stability of IRV drug substance (DS) and aluminum-formulated drug product (DP) when stored at -70±10°C and 5±3 °C, respectively. Our results support the use of this EIA to examine the stability and determine the potency, antigen dose, lot-to-lot consistency, and lot release of IRV products. This RV potency assay may serve as an alternative to in vivo potency tests, making it suitable for quality control tests of cGMP IRV lots in clinical trials.
为了确定灭活轮状病毒疫苗(IRV)的效价,我们开发了一种酶免疫测定法(EIA),该方法使用生物素偶联的RV VP7特异性单克隆抗体。RV VP7是外衣壳层中的一种关键结构蛋白,决定RV G基因型并引发宿主免疫反应,包括中和抗体。这种EIA具有高特异性、良好的线性、高精度和高准确性,RV抗原的检测下限(LOD)和定量下限(LOQ)低至0.037µg/ml。分别在-70±10°C和5±3°C储存时,对该EIA进行了评估,结果证明其适用于确定IRV原料药(DS)和铝盐制剂药品(DP)的长期稳定性。我们的结果支持使用这种EIA来检测IRV产品的稳定性,并确定其效价、抗原剂量、批次间一致性和批次放行情况。这种RV效价测定法可替代体内效价试验,适用于临床试验中cGMP IRV批次的质量控制检测。