中文摘要：

目的：建立Biacore法测定重组人乳头瘤病毒双价（16/18型）疫苗原液的动力学常数。方法：分别将HPV 16·V5中和单抗与HPV 18·J4中和单抗偶联到CM5芯片上，然后根据Biacore X-100 Kinetics/Affinity控制软件将 HPV 16L1和HPV 18L1原液分别稀释至适当的不同浓度，采用50 mmol·L^-1 氢氧化钠溶液作为再生液进行动力学分析。根据Biacore X-100 Evaluation分析软件进行拟合，得到样品的动力学常数KD值。结果：Biacore法测定重组人乳头瘤病毒16型、18型原液动力学常数KD分别为4.844×10^-10 mol·L^-1和1.67×10^-10 mol·L^-1，HPV 16L1和HPV 18L1原液在浓度为0.625-40 μg·mL^-1时线性关系良好（HPV16L1：Y=17.77X-0.0184，r=1.000；HPV 18L1：Y=18.73X＋5.282，r=0.9957）。HPV 16L1和HPV 18L1原液精密度和稳定性试验的RSD均〈20%。并且Biacore法测定的结合常数ka值和体外相对效力测定（IVRP法）的EC50值之间具有一定的相关性。结论：该方法实时、简便、快速、准确、灵敏，适用于重组人乳头瘤病毒原液的动力学分析。

英文摘要：

Objective： To establish a surface plasmon resonance biosensor （Biacore X-100） to detect the two KD values of recombinant human papillomavirus types 16 and 18 bulks. Methods： HPV 16·V5 and HPV 18·J4 neutralizing monoclonal antibodies were coupled to sensor chip CM5,respectively.With the help of the control software of Biacore X-100 Kinetics/Affinity,HPV 16 L1 and HPV 18L1 bulks were diluted to several suitable concentrations respectively and 50 mmol·L^-1 NaOH was used as a regeneration solution for kinetic analysis.The data was analyzed to calculate the KD values with fitting process by Biacore X-100 evaluation software. Results： The KDvalues of HPV 16 and HPV 18 bulks by Biacore were 4.844×10^-10 mol·L^-1 and 1.67×10^-10 mol·L^-1,respectively.The linear concentration of original HPV 16L1 and HPV 18L1 solutions ranged from 0.325 to 160 μg·mL^-1（for HPV16L1,Y =17.77X-0.0184,r=1.000;for HPV 18L1,Y =18.73X＋5.282,r=0.9957）.The RSD of precision and stability study was below 20%.The ka value detected by Biacore had certain relevance to the EC50 values of IVRP. Conclusion： The method is real-time,simple,rapid,accurate and sensitive.It can be used to detect the kinetic constants of HPV bulks.