久久久精品欧美一区二区三区,国产欧美久久久精品影院,亚洲AV福利天堂一区二区三亚洲欧洲日韩国产AⅤ在线,亚洲成av人在线视,四虎国产精品永久地址99自拍三区一级国产片,亚洲男人的天堂久久无在线观看免费黄视频,亚洲一级特黄大片在线播放,国产成人精品区在线观看

Gene Therapy
Home > Application > Gene Therapy > Adeno-Associated Virus

Adeno-Associated Virus

Adeno-associated virus (AAV) is the most commonly used vector in gene therapy, with a diameter of about 20 nm and no envelope. Its genome is about 4.8 kb in length. AAV does not encode DNA polymerase and relies on host cells or helper viruses to synthesize its own genome, so it cannot replicate autonomously. AAV is quite prevalent in humans and other primates, with 11 known serotypes. In addition, many artificial variants of AAV have been developed to improve the efficiency of AAV as vectors in clinical and research applications. For example, Glybera?, the world's first approved AAV-based product, uses AAV1 to deliver lipoprotein lipase (LPL) to treat patients with LPL deficiency. Luxturna? was approved in 2017 as an AAV2-based treatment for Leber's congenital amaukia, which causes progressive blindness, and Zolgensma? was approved in 2019 as an AAV9-based treatment for spinal muscular atrophy (SMA).

 

There are different platform strategies for the upstream production of AAV, including herpes simplex virus (HSV) infection, insect cell-baculovirus expression vector system, and stable production cell line, but the method based on transient transfection of HEK293 cell by plasmid DNA is currently the mainstream method for the production of clinical materials, which is more flexible and easy to use, with low up-front engineering effort, thus can help to quickly initiate the first-in-human studies. Typical process steps for the production of AAV based on transient transfection include: adherent or suspension culture based cell expansion, plasmid transfection, viral vector production, cell lysis, purification – including clarification, affinity capture chromatography, ion exchange polish chromatography, tangential flow filtration for concentration/diafiltration, final sterile filtration and fill/finish.

 

Adeno-Associated Virus

 

The choice of adherent vs. suspension culture should be comprehensively considered based on the technical capabilities of the platform, the stages, target indications and administration routes, and the expected demand for viral vectors. For large patient population , applications that require systemic administration or higher levels of viral vectors per dose, developing a production strategy based on suspension culture will be more conducive to ensuring the required scale-up capability, and it has been reported that AAV suspension culture production up to 2,000 L has been achieved. The purpose of upstream process development is to explore the optimal conditions for virus production, including improving transfection efficiency and ensuring its consistency under large-scale production conditions by adjusting transfection conditions, and intensifying virus productivity through strategies such as optimizing media formulations, maintain viral infectivity.

 

Although some AAV serotypes will partially secrete virus particles into the cell culture media during the culture process, in order to increase the yield of the virus, the downstream process of AAV starts with mechanical or chemical lysis of cells and DNA fragmentation under closed conditions, Subsequent clarification steps are designed to remove residual large amounts of plasmid DNA as well as host cell contaminants from cell lysis. Filter selection should consider the level of turbidity reduction, high product yield, and ease of scale-up. For the capture of AAV by affinity chromatography, there are currently specific affinity resin for various serotypes. Usually, a single-step affinity capture can achieve high purity. However, in order to ensure the specific level of impurity removal, ion exchange chromatography is generally used in combination as a polishing step. Especially for the challenge of separating empty and full capsids in AAV production, ion-exchange chromatography is currently the most commonly used method at production scale. Empty capsids are considered as in-process impurity by regulators and their percentage needs to be reduced as much as possible and the empty/full capsid ratio controlled to ensure consistency of gene therapy products. Tangential flow filtration aims to concentrate the product and replace it with a suitable buffer system. In view of the small particle size of AAV, MWCO 100kD membrane modules can be selected, while sterile filtration aims to reduce microbial load and meet the required sterility regulatory requirements.


Related Products
Contact Us
  • Tel.: +86 021 6434 0155
  • E-Mail: marketing@duoningbio.com
  • Address: Building 30, No. 1525 Minqiang Road, Songjiang District, Shanghai
Message

Copyright ? Shanghai Duoning Biotechnology Co., Ltd. All Rights Reserved Sitemap | Technical Support: Reanod

WeChat
WeChat

Message-

Duoning Biotechnology

+86 021 6434 0155

美丽人妻无套中出中文字幕 | 久久九九有精品国产| 色窝窝无码一区二区三区| 少妇厨房愉情理伦BD在线观看| 狠狠色丁香久久婷婷综合五月| 亚洲日本VA午夜在线电影| 男生被男人CAO屁股的后果| 无码专区久久综合久综合字幕| 久久无码喷吹高潮播放不卡| 少妇高潮喷水久久久影院| 曰批视频免费40分钟野战| 中文成人无字幕乱码精品| 国内精品久久久久久99蜜桃| 国产亚洲欧美另类精品久久久| 欧美自拍嘿咻内射在线观看| 成人H动漫精品一区二区无码| 不卡AV片在线看| 成人免费777777被爆出| 噜噜噜亚洲色成人网站∨| 成人毛片无码一区二区三区| 毛多水多高潮高清视频| 色悠悠久久久综合88| 国产成人亚洲综合一区| 高清中文字幕男人的天堂| 国产成人无码精品一区在线观看| 精品国产鲁一鲁一区二区交| 无码免费一区二区三区免费播放| 岛国少妇精品久久中文字幕| 日韩欧美精品一区二区三区视频在线观看| 国产精品一区二区尿失禁| 在线天堂中文最新版网| 男人的天堂欧美网站免费观看欧美日韩ay在线观看| 国产女主播精品大秀系列| 国产99视频精品专区| 久久久久亚洲Av专区首页| 人人妻人人爽人人添夜夜欢视频| 亚欧免费无码Aⅴ在线观看网站| 日本卡一卡二不卡| 国产成人精品日本亚洲黑人| 99久久精品无码一区二区毛片免费| 成人精品视频在线观看亚洲超碰|