枢密增强版AAV-PHP.eB !

       枢密研发团队目前已有成功高效产品：AAV-PHP.eB，测试结果见文末。

图2 .外周及中枢神经系统转导效果

        为了获得高转导效率，科学实验中已经进行了许多努力如使用梯度离心或柱色谱法纯化AAV载体。但这些方法耗时长，成本高并且需要昂贵的设备^[6]。

        枢密科技大规模AAV制备系统，有效解决了这些问题，我司采用杆状病毒-SF9细胞包装系统，联合无血清悬浮培养生物反应器可进行大规模发酵，极大地提高产量，进而满足大规模工业化制备的要求。

枢密测试结果 
 

图注：尾静脉注射PHP.B血清型的rAAV-hSyn-mCherry-WPRE-pA，可看到病毒可有效跨越BBB，高效标记小鼠大脑CNS。图A所示为小鼠脑矢状切片，图B、C和D为A虚线框的放大图。
 

图注：尾静脉注射PHP.eB血清型的rAAV-CamKIIα-EYFP-WPRE-pA，可看到病毒可有效跨越BBB，高效标记小鼠大脑CNS。

参考文献

1. Mathiesen SN, Lock JL, Schoderboeck L, Abraham WC, Hughes SM.CNS Transduction Benefits of AAV-PHP.eB over AAV9 Are Dependent on Administration Route and Mouse Strain. Mol Ther Methods Clin Dev.2020;19:447-458.

2. Xie BS, Wang X, Pan YH, Jiang G, Feng JF, Lin Y. Apolipoprotein E, low-density lipoprotein receptor, and immune cells control blood-brain barrier penetration by AAV-PHP.eB in mice.Theranostics.2021;11(3):1177-1191.

3. Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sánchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat Neurosci.2017;20(8):1172-1179.

4. Hu S, Yang T, Wang Y. Widespread labeling and genomic editing of the fetal central nervous system by in utero CRISPR AAV9-PHP.eB administration. Development. 2021;20;148(2).

5. Luoni M, Giannelli S, Indrigo MT, Niro A, Massimino L, Iannielli A, Passeri L, Russo F, Morabito G, Calamita P, Gregori S, Deverman B, Broccoli V. Whole brain delivery of an instability-prone Mecp2 transgene improves behavioral and molecular pathological defects in mouse models of Rett syndrome. Elife. 2020 Mar;24;9:e52629.

6. Konno A, Hirai H.Efficient whole brain transduction by systemic infusion of minimally purified AAV-PHP.eB.J Neurosci Methods.2020;346:108914.