Shank-Retzlaff Mary L, Zhao Qinjian, Anderson Carrie, Hamm Melissa, High Katrina, Nguyen Mai, Wang Feng, Wang Ning, Wang Bei, Wang Yang, Washabaugh Michael, Sitrin Robert, Shi Li
Merck Research Laboratories Department of Bioprocess, West Point, Pennsylvania, USA.
Hum Vaccin. 2006 Jul-Aug;2(4):147-54. doi: 10.4161/hv.2.4.2989. Epub 2006 Jul 4.
The thermostability of GARDASIL (Merck & Co., Inc, Whitehouse Station, NJ, USA), a developmental vaccine against human papillomavirus (HPV), was evaluated using an enzyme immunoassay, referred to as the in vitro relative potency (IVRP) assay and differential scanning calorimetry (DSC). Gardasil samples were stored at temperatures ranging from 4 to 42 degrees C and tested for IVRP at various time points. Extrapolation of the IVRP results indicates GARDASIL is extremely stable. The half-life of the vaccine is estimated to be 130 months or longer at temperatures up to 25 degrees C. At 37 degrees C, the half-life is predicted to be 18 months and at 42 degrees C, the half-life is predicted to be approximately three months. Differential scanning calorimetry (DSC) analysis was used to evaluate the process of protein denaturation during a rapid temperature increase (as opposed to long-term storage at a specific temperature). Differences were seen among the DSC profiles of the four HPV types tested. This indicates that small differences in the amino acid structure can have a significant effect on the intermolecular contacts that stabilize the L1 proteins and the VLP assembly. For the Gardasil samples evaluated here, DSC results demonstrated the relative overall structural stability of the VLPs, but were not predictive of the excellent long-term stability observed with the IVRP assay.
利用一种酶免疫测定法（即体外相对效价（IVRP）测定法）和差示扫描量热法（DSC），对一种处于研发阶段的抗人乳头瘤病毒（HPV）疫苗佳达修（美国新泽西州怀特豪斯站默克公司生产）的热稳定性进行了评估。将佳达修样品储存在4至42摄氏度的温度范围内，并在不同时间点进行IVRP测试。IVRP结果的外推表明佳达修极其稳定。在高达25摄氏度的温度下，该疫苗的半衰期估计为130个月或更长。在37摄氏度时，半衰期预计为18个月，在42摄氏度时，半衰期预计约为三个月。差示扫描量热法（DSC）分析用于评估在温度快速升高过程中（与在特定温度下长期储存相反）蛋白质变性的过程。在所测试的四种HPV类型的DSC图谱中观察到了差异。这表明氨基酸结构上的微小差异会对稳定L1蛋白和病毒样颗粒（VLP）组装的分子间接触产生重大影响。对于此处评估的佳达修样品，DSC结果证明了VLP相对整体的结构稳定性，但无法预测IVRP测定法所观察到的出色的长期稳定性。
