重组腺相关病毒基因药物三种滴度的比较与分析

采用酶联免疫吸附测定(ELISA)、实时荧光定量聚合酶链反应(Q-PCR)和转导方法对重组腺相关病毒(rAAV)的衣壳滴度、基因组滴度、转导滴度进行定量,并比较衣壳滴度/基因组滴度(P/GC)和基因组滴度/转导滴度(GC/TU).实验结果表明:3批rAAV的平均衣壳滴度为3.65×10¹³ P·mL^-1,平均基因组滴度为8.67×10¹¹ GC·mL^-1,平均转导滴度为9.85×10⁹ TU·mL^-1,P/GC平均值为41.70,GC/TU平均值为88.20,P/GC和GC/TU平均值接近于AAV2标准品,说明文中方法的制备工艺成熟、稳定,制备的rAAV感染活性较高.

Comparison and Analysis of Three Titers of Recombinant Adeno-Associated Virus Gene Drugs

The particle titer, genome titer and transduction titer of recombinant adeno-associated virus(rAAV)were quantified by enzyme-linked immuno sorbent assay(ELISA), quantitative real-time polymerase chain reaction(Q-PCR)and transduction method, and then the particle titer/genome titer(P/GC)and genome titer/transduction titer(GC/TU)were compared. The experiment results showed that the average particle titer of rAAV in three batches was 3.65×10¹³ P·mL^-1, the average genome titer was 8.67×10¹¹ GC·mL^-1, the average transduction titer was 9.85×10⁹ TU·mL^-1, the average value of P/GC was 41.70 and the average value of GC/TU was 88.20. The average values of P/GC and GC/TU were close to the AAV2 standard, indicating that the preparation process of the method in this paper is mature and stable, and the prepared rAAV has high infectious activity.