Objective: To analyze the particle titer and full rate of a recombinant adeno-associated virus (rAAV) by using size exclusion chromatography (SEC) with tandem ultraviolet detector (UV), multi angle laser scattering detector (MALS) and differential detector (RI). Methods: SEC-UV-MALS-RI series analysis was performed on an rAAV, its empty shell reference substance, and a mixture of the two samples. The mobile phase was 50 mmol·L-1 phosphate buffer, with a flow rate of 0.3 mL·min-1, and a sample loading volume of 50 μL. The temperature is room temperature and the collection time was 30 min. The UV detector selected dual channels of 260 nm and 280 nm. 18 angle laser scattering detection and differential detection adopted the system default settings. The results were analyzed using WYATT software. Results: The particle titer of the sample was 4.816×1012 vp·mL-1, with an RSD of 0.37%. The particle titer of the empty shell reference substance was 3.199×1012 vp·mL-1, with an RSD of 0.75%. The particle titer RSD of the 5 samples mixed according to the preset different ratios were all within 3%, and the RSD of full ratio was all within 4%. The measurement results of particle titer and full ratio of the mixed sample were basically consistent with the expected values. Conclusion: The SEC-UV-MALS-RI method can be used to analyze the particle titer, full ratio, and other items of rAAV products, which is suitable for quality control of particle titer and full ratio of rAAV products.
LI Xiang, SHI Xin-chang, WANG heng, HAN Chun-mei, QIN Xi, ZHOU Yong
. Analysis of particle titer and full ratio of a recombinant adeno-associated virus (rAAV) by SEC-UV-MALS-RI technique*[J]. Chinese Journal of Pharmaceutical Analysis, 2023
, 43(11)
: 1840
-1846
.
DOI: 10.16155/j.0254-1793.2023.11.05
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