Shank-Retzlaff M, Wang F, Morley T, Anderson C, Hamm M, Brown M, Rowland K, Pancari G, Zorman J, Lowe R, Schultz L, Teyral J, Capen R, Oswald C B, Wang Y, Washabaugh M, Jansen K, Sitrin R
Merck Research Laboratories Department of Bioprocess & Bioanalytical Research, West Point, Pennsylvania, USA.
Hum Vaccin. 2005 Sep-Oct;1(5):191-7. doi: 10.4161/hv.1.5.2126. Epub 2005 Sep 20.
An in vitro relative potency (IVRP) assay has been developed as an alternative to the mouse potency assay used to release Merck's human papillomavirus (HPV) vaccine, Gardasil, for early phase clinical trials. The mouse potency assay is a classical, in vivo assay, which requires 4-6 weeks to complete and exhibits variability on the order of 40% relative standard deviation (RSD). The IVRP assay is a sandwich-type immunoassay that is used to measure relative antigenicity of the vaccine product. The IVRP assay can be completed in three days, has a variability of approximately 10% RSD and does not require the sacrifice of live animals. Because antigen detection is achieved using H16.V5, a neutralizing monoclonal antibody, which binds to a clinically-relevant epitope, the relative antigenicity measured by the IVRP assay is believed to be a good predictor of in vivo potency. In this study, the relationship between immunogenicity, as measured by the mouse potency assay and antigenicity as measured by the IVRP assay, is demonstrated. Freshly manufactured and aged samples produced using two different manufacturing processes were tested using both methods. The results demonstrate that there is an inverse correlation between the IVRP and mouse potency assays. Additionally, clinical results indicate IVRP is predictive of human immunogenicity. Thus, antigenicity, as defined by the H16.V5 epitope, can be used as a surrogate for immunogenicity and the IVRP assay is suitable for use as the sole potency test for Gardasil samples.
已开发出一种体外相对效价（IVRP）测定法，作为默克公司用于人乳头瘤病毒（HPV）疫苗佳达修早期临床试验放行的小鼠效价测定法的替代方法。小鼠效价测定法是一种经典的体内测定法，需要4至6周才能完成，相对标准偏差（RSD）约为40%。IVRP测定法是一种夹心型免疫测定法，用于测量疫苗产品的相对抗原性。IVRP测定法可在三天内完成，RSD约为10%，且无需处死实验动物。由于使用与临床相关表位结合的中和单克隆抗体H16.V5进行抗原检测，因此IVRP测定法测得的相对抗原性被认为是体内效价的良好预测指标。在本研究中，展示了通过小鼠效价测定法测得的免疫原性与通过IVRP测定法测得的抗原性之间的关系。使用两种不同生产工艺生产的新鲜制备和老化样品均采用这两种方法进行了检测。结果表明，IVRP测定法与小鼠效价测定法之间存在负相关。此外，临床结果表明IVRP可预测人体免疫原性。因此，由H16.V5表位定义的抗原性可作为免疫原性的替代指标，且IVRP测定法适合用作佳达修样品的唯一效价检测方法。
