AAV 캡시드 엔지니어링

장점

품질

• Excellent Input Library Quality

  

  Excellent pre-screening library diversity and uniformity

   

  A high coverage library contains a large number of distinct capsid variants, which increases the likelihood of finding variants with desirable properties. A low coverage library may miss important capsid variants, limiting the potential for capsid engineering. Therefore, it is important to ensure that the library is diverse and contains a large number of distinct variants.​

  Uniformity is also important because it ensures that each capsid variant is represented equally in the library. Uneven representation can bias the screening process and lead to the selection of suboptimal variants. Therefore, it is important to ensure that the library is evenly distributed and that each variant is represented in similar amounts.​

   

  

  Extremely low mispackaging rate : <0.1%

   

  Percentage of virus packed with early terminated Capsid gene is a key indicator of mispackaging rate. High mispackage rate will lead to mismatch of the capsid serotype with the actual genome it contains, resulting in error in the screening result.

  In AAV capsid engineering, the goal is to modify the AAV capsid to improve its efficiency or specificity. One way to achieve this is to generate a library of AAV variants and screen them for desired properties. However, if the AAV vectors produced from this library have a high mispackaging rate, the screening results may be inaccurate and unreliable.

  A high mispackaging rate can result in the production of AAV vectors that contain incorrect genetic material. These vectors can then transduce unintended cells or tissues, potentially causing toxicity or reduced efficacy. Additionally, high levels of mispackaging can lead to the formation of replication-competent AAV (rcAAV) particles, which can cause adverse immune reactions and limit the safety of therapies for cancer or genetic disorder.

  Therefore, it is important to minimize the mispackaging rate in AAV capsid engineering. This can be achieved by optimizing the vector production process, such as controlling the input DNA amount, using high-quality plasmids, and optimizing the transfection conditions. Additionally, careful screening of the AAV vectors for mispackaging should be performed to ensure that the vectors being tested are representative of the intended capsid library.

  To measure mispackaging rate, we measure the rate of AAV population with early terminated capsid protein in the total population. The result indicate we have extremely low mispackaging rate , ensuring the matching of packed AAV capsid gene and the capsid protein during virus packaging, thus lead to reliable screening results.

• Novel Capsid screened from π-Icosa system target to CNS

  

• Novel capsid targeting muscle with liver-detargeting

  PG007- Novel capsid targeting muscle​

  

   

  Mouse samples are collected 21 days after virus injection
  AAV-PG007, a modified backbone via π-Icosa system based on AAV9 and AAV2 chimera shows a reduction of the off-targeting to the liver while keeping muscle targeting in both mouse and monkey​

  

  PG007 is both efficient and specific in muscle-targeting cross rodent species​

• Novel capsid targeting primary human T cells

  Infection of Quiescent Primary Human T Cells with Novel PackGene AAV6 Serotypes -- Imaging ​

  • 4 novel capsid: PG010, PG012, PG013, PG014​
  • 10x MOI compared to WT​

  Infection of Quiescent Primary Human T Cells with Novel PackGene AAV6 Serotypes – FACS Analysis​

  

  • 4 novel capsid: PG010, PG012, PG013, PG014​
  • 10x MOI compared to WT​

  Infection of Quiescent Primary Human T Cells with Novel PackGene AAV6 Serotypes – mRNA and luciferase ​

  

자료

• Generation of novel AAV serotype with enhanced infectivity, specificity, and lower toxicity via π-Icosa AAV Capsid Engineering platform

• AAV Capsid Engineering Service Flyer

• Capsid Engineering Application Note

• Capsid Engineering Nature Editorial