HBV rAAV載體的特點

我們的優勢

技術說明

• 操作要求

  rAAV作為基因治療載體的安全性在1994年被FDA認可，其生物安全性為一級（BSL-1），與質粒DNA的安全級別相同，建議使用BSL-2實驗室和ClassⅡ生物安全柜。

• 儲存要求

  • 將病毒儲存在 -80°C冰箱， 并在操作過程中將其置于冰上。
  • 建議對使用量提前進行預估，派真生物出貨時按照預定的分裝要求進行分裝，避免收到貨后再次凍融分裝影響滴度準確性。如需再次分裝使用，推薦使用內壁硅化程度良好的PCR管，或者低蛋白結合率的病毒專用保存管。
  • 如需稀釋AAV-HBV，請將病毒置于冰浴融解后，使用Tris緩沖液稀釋。

• AAV-HBV小鼠造模實驗設計

  重組腺相關病毒（rAAV）已經被廣泛應用于動物體內實驗和細胞研究中。但對于體內試驗，注射部位如何選擇，注射多少病毒量，注射后多久檢測等問題，因為研究領域、研究的動物不同，而很難有簡單通用的注射方法，實驗中還需要具體探索最佳條件：

  強烈建議在正式實驗之前，對AAV-HBV進行3~4個注射量的梯度測試，每只小鼠依次按最低要求遞增。比如：1E+11 GC；2E+11 GC；3E+11 GC 做3個梯度稀釋；

• 客戶案例

  新型抗HBV治療方法的研究

  客戶發表文章：Meng C, Sun S, Liang Y, et al. Engineered anti-PDL1 with IFNαtargets both immunoinhibitory and activating signals in the liver to break HBV immune tolerance. Gut 2023; 72: 1544-1554.

  病毒類型：rAAV8-HBV ayw，D基因型

  注射方式：C57BL/6小鼠，尾靜脈注射

  注射劑量：1 E +11 GC

  檢測時間：35天

  

• 相關文獻

  1. Gao L, Yang J, Feng J, Liu Z, Dong Y, Luo J, Yu L, Wang J, Fan H, Ma W and Liu T (2022)PreS/2-21-Guided siRNA Nanoparticles Target to Inhibit Hepatitis B Virus Infection and Replication. Front. Immunol. 13:856463. doi: 10.3389/fimmu.2022.856463.
  2. Jing Miao, Xiqin Yang, Xuwei Shang, Zhe Gao, Qian Li, Yun Hong, Jiaying Wu, Tingting Meng, Hong Yuan, Fuqiang Hu. Hepatocyte-targeting and microenvironmentally responsive glycolipid-like polymer micelles for gene therapy of hepatitis B. Molecular Therapy - Nucleic Acids,Volume 24, 2021, Pages 127-139.
  3. Xu W, Niu Y, Ai X, Xia C, Geng P, Zhu H, Zhou W, Huang H, Shi X. Liver-Targeted Nanoparticles Facilitate the Bioavailability and Anti-HBV Efficacy of Baicalin In Vitro and In Vivo. Biomedicines. 2022; 10(4):900.
  4.Wang X, Zhang Y, Ben Y, Qiu C, Wu J, Zhang W, Wan Y. Anti-HBc IgG Responses Occurring at the Early Phase of Infection Correlate Negatively with HBV Replication in a Mouse Model. Viruses. 2022; 14(9):2011.
  5. Yue Wang, Yaxian Mei, Zhenghong Ao, Yuanzhi Chen, Yichao Jiang, Xiaoqing Chen, Ruoyao Qi, Baorong Fu, Jixian Tang, Mujin Fang, Min You, Tianying Zhang, Quan Yuan, Wenxin Luo, Ningshao Xia. A broad-spectrum nanobody targeting the C-terminus of the hepatitis B surface antigen for chronic hepatitis B infection therapy. Antiviral Research, Volume 199, 2022, 105265.
  6. Xiu-Qing Pang, Xing Li, Wei-Hang Zhu, Run-Kai Huang, Zhi-shuo Mo, Ze-Xuan Huang, Yuan Zhang, Dong-Ying Xie, Zhi-Liang Gao. LAG3+ erythroid progenitor cells inhibit HBsAg seroclearance during finite pegylated interferon treatment through LAG3 and TGF-β. Antiviral Research, Volume 213, 2023, 105592.
  7. Meng C, Sun S, Liang Y, et al. Engineered anti-PDL1 with IFNα targets both immunoinhibitory and activating signals in the liver to break HBV immune tolerance. Gut 2023; 72: 1544-1554.
  8. Y. Jiang, X. Chen, X. Ye, C. Wen, T. Xu, C. Yu, W. Ning, G. Wang, X. Xiang, X. Liu, Y. Wang, Y. Chen, X. Liu, C. Shi, C. Liu, Q. Yuan, Y. Chen, T. Zhang, W. Luo, N. Xia. A Dual-domain Engineered Antibody for Efficient HBV Suppression and Immune Responses Restoration. Adv. Sci. 2024, 11, 2305316.
  9. Wang, Y., Guo, L., Shi, J. et al. Interferon stimulated immune profile changes in a? humanized mouse model of HBV infection. Nat Commun 14, 7393 (2023).