Adenovirus Amplification and purification Service
Begin your customized ADV production process
Introduction to Adenovirus Amplification and purification Service
The adenoviral vectors provided by Genemedi are based on human adenovirus type 5 (Ad5), which is replication-incompetent (-E1/-E3) and can’t be integrated into host genome, guaranteeing the security for subsequent operations. Adv packaging by Genemedi shows almost 100% gene delivery in most cell types in the recombinant protein expression system both in vivo and in vitro. Recombinant protein expression of interest is detectable from 24 hours post infection. With the premade adenovirus bought from Genemedi or provided by customers, Genemedi launchs high quality adenovirus amplification service. Adenovirus amplified by Genemedi will be tested to guarantee the quality, the titer of which can be up to 1*10^11 PFU/ml.
|Sipping and Storage Guidelines||Shipped by dry ice, stored at -80 ° C, effective for 1 year. Avoid repeatedly freezing and thawing|
|Titer||> 1*10^11 PFU/ml|
1. Broad range of host. Adv has the ability to infect dividing, quiescent cells, stem cells, and primary cells, allowing genetic materials to be delivered to a highly diverse range of cell types and tissues.
2. High infection efficiency. Almost100% gene delivery in most cell types, completely surpassing other viral vector tools and liposome transfection.
3. High titer. Up to 1011 pfu/ml.
Applications and Figures
Quality control description
Our optimized custom adenoviral vector production and strict quality control systems provide customers a high titer of functional recombinant adenoviral vectors. Viral titers are determined by TCID50 method, which is the most accurate way to measure the titer of adenovirus.
1. For detailed protocol about how to package and purify Adv, please see the Adenovirus User Manual on the Genemedi website.
Frequently Asked Questions(FAQs)
- 1. Why does adenovirus have a relatively higher immunogenecity compared with rAAV?
The adenovirus without E1/E3 can express all of the other genes in the viral backbone and hence induces immunogenic responses, while rAAV does not have any of the AAV genes, thus no immunogenicity from viral protein.
- 2. What is the role of the E1 Gene in adenoviruses?
Simply, the E1 gene products are early proteins that are transcribed in the early transcribed regions and required for proceeding subsequent steps in viral replication. The E1 gene contains E1A and E1B, involved in the replication of adenovirus. E1A is critical to start viral replication by promoting transcription from rep gene promoters, P5 and P19, and facilitate viral replication by activating the early adenovirus promoters.
- 3. How to store adenovirus?
It would be better to store adenovirus in PBS at -80oC. Sucrose or DMSO may help to stabilize the vector.
- 4. How can you tell if your vector is lentiviral, retroviral, or adenoviral?
You should blast your vector sequence and see if there’re sequences of reverse transcriptase and integrase (gene names: gag and pol), which are for lentiviral/retroviral vectors, but not for adenoviral. For another, if your plasmid is around 30-35kb in size, it's certainly adenoviral.
- 5. Is adenovirus a useful tool to study primary macrophage functions?
In RAW264.7 and PM cells, adenovirus works very well, and it seems that IL1β expression is increased slightly after adenovirus transfection compared with negative control. While for the BMDM, adenovirus does not work, it may be better to use lentivirus instead, which gives a pretty good transduction efficacy and less inflammatory response.
- 6. Is it possible to infect a tissue preparation with lentivirus and afterwards with adenovirus and getting high efficiency in transduction?
It is definitively possible to perform sequential transductions/infections. Polybreen, protamine sulfate or other transduction enhancing reagents are recommended to enhance viral particle infectivity.
1. Hu J, L Zhang, Y Yang, Y Guo, Y Fan, M Zhang, W Man, E Gao, W Hu, RJ Reiter, H Wang and D Sun. (2017). Melatonin alleviates postinfarction cardiac remodeling and dysfunction by inhibiting Mst1. J Pineal Res 62.
2. Xu C, F Wu, C Mao, X Wang, T Zheng, L Bu, X Mou, Y Zhou, G Yuan, S Wang and Y Xiao. (2016). Excess iodine promotes apoptosis of thyroid follicular epithelial cells by inducing autophagy suppression and is associated with Hashimoto thyroiditis disease. J Autoimmun 75:50-57.
3. Wang Y, X Zhao, X Wu, Y Dai, P Chen and L Xie. (2016). microRNA-182 Mediates Sirt1-Induced Diabetic Corneal Nerve Regeneration. Diabetes 65:2020-31.
4. Luo T, J Fu, A Xu, B Su, Y Ren, N Li, J Zhu, X Zhao, R Dai, J Cao, B Wang, W Qin, J Jiang, J Li, M Wu, G Feng, Y Chen and H Wang. (2016). PSMD10/gankyrin induces autophagy to promote tumor progression through cytoplasmic interaction with ATG7 and nuclear transactivation of ATG7 expression. Autophagy 12:1355-71.
5. Liu TY, XQ Xiong, XS Ren, MX Zhao, CX Shi, JJ Wang, YB Zhou, F Zhang, Y Han, XY Gao, Q Chen, YH Li, YM Kang and GQ Zhu. (2016). FNDC5 Alleviates Hepatosteatosis by Restoring AMPK/mTOR-Mediated Autophagy, Fatty Acid Oxidation, and Lipogenesis in Mice. Diabetes 65:3262-3275.
6. Liu H, S Fang, W Wang, Y Cheng, Y Zhang, H Liao, H Yao and J Chao. (2016). Macrophage-derived MCPIP1 mediates silica-induced pulmonary fibrosis via autophagy. Part Fibre Toxicol 13:55.
7. Yao T, X Ying, Y Zhao, A Yuan, Q He, H Tong, S Ding, J Liu, X Peng, E Gao, J Pu and B He. (2015). Vitamin D receptor activation protects against myocardial reperfusion injury through inhibition of apoptosis and modulation of autophagy. Antioxid Redox Signal 22:633-50.
8. Wang B, A Ma, L Zhang, WL Jin, Y Qian, G Xu, B Qiu, Z Yang, Y Liu, Q Xia and Y Liu. (2015). POH1 deubiquitylates and stabilizes E2F1 to promote tumour formation. Nat Commun 6:8704.
9. Hua F, K Li, JJ Yu, XX Lv, J Yan, XW Zhang, W Sun, H Lin, S Shang, F Wang, B Cui, R Mu, B Huang, JD Jiang and ZW Hu. (2015). TRB3 links insulin/IGF to tumour promotion by interacting with p62 and impeding autophagic/proteasomal degradations. Nat Commun 6:7951.
10. Wang X, J Liu, J Zhen, C Zhang, Q Wan, G Liu, X Wei, Y Zhang, Z Wang, H Han, H Xu, C Bao, Z Song, X Zhang, N Li and F Yi. (2014). Histone deacetylase 4 selectively contributes to podocyte injury in diabetic nephropathy. Kidney Int 86:712-25.