project background
Adeno-associated virus (AAV) is one of the most widely used viral vectors in the field of gene therapy at present. Due to its advantages such as high safety, long-term stable expression and low immunogenicity, it has become the preferred delivery tool for the treatment of genetic diseases, neurodegenerative diseases, ophthalmic diseases, etc. AAV is a filamentous DNA virus belonging to the Parvoviridae family without an envelope. It can only replicate in the presence of helper viruses (usually adenovirus, herpes simplex virus, etc.).
Recombinant adeno-associated virus (rAAV) is a gene vector modified on the basis of wild-type AAV. It was modified by deleting the rep and cap genes of wild-type AAV, retaining only the reverse terminal repeat sequences (ITRs), and inserting therapeutic gene expression kits into them.
Replicating adeno-associated virus (ReplicationCompetentAAV, rcAAV) is an AAV with replicating ability formed during the production of rAAV vectors due to non-homologous recombination of the rep/cap gene and ITR. Although rcAAV particles have no direct association with the currently known human diseases, as contaminants, they can enhance or interfere with the therapeutic effect of rAAV or cause the drift of rAAV vector gene sequences, affecting their genomic stability and in vivo transduction efficiency, etc. Therefore, it is of great significance to monitor rcAAV during large-scale production and to test it when products are released.
Regulatory requirements
The "Technical Guidelines for Pharmaceutical Research and Evaluation of In Vivo Gene Therapy Products (Trial)" issued by the Center for Drug Evaluation (CDE) of the National Medical Products Administration emphasizes the establishment of a complete quality control strategy based on quality research and the identification of key quality attributes. In the process of virus vector preparation, control strategies should be established to address the potential risks of replicable viruses, such as conducting replicable virus detection on samples or final products at appropriate stages of the production process.
In the "Technical Guidelines for Non-clinical Research and Evaluation of Gene Therapy Products", it is mentioned that for gene therapy products using replication-defective viral vectors, it is necessary to assess whether there is a possibility of generating replicative viruses during the production process.
Product features
The Baorui Biotech RCAAV-2 /N Detection Kit (PCR-fluorescence probe method) consists of two systems: the system for detecting rcAAV and the system for detecting rAAV. The calibrators of the kit are ready-to-use calibrators that have been calibrated by digital PCR technology and do not require dilution before use. They can simultaneously achieve rapid, sensitive and specific quantitative detection of rcAAV and rAAV.
This kit has the following features:
01.
Good precision
Repeatability (intra-analytical precision), intermediate precision (inter-instrument, inter-day, inter-personnel, inter-reagent batch) and reproducibility (total imprecision), the quantification limit CV≤20%, and the CV≤15% for medium and high concentrations.
02.
Strong specificity
It has no cross-reaction with commonly used cell lines such as HEK293, Sf9, Hi5, CHO, Escherichia coli, Pichia pastoris, and Vero. There is no cross-reaction with Staphylococcus aureus, Staphylococcus epidermidis, mycobacteria, etc.
03.
Convenient and fast operation
1. Single-tube PCR reagents, easy to access, no need to prepare reagents when in use;
2. Calibrators do not need to be diluted, and the finished products can be assembled conveniently and quickly.
From sample extraction to the completion of the test, it only takes about 1.5 hours.
04.
Anti-pollution design
The UDG enzyme anti-contamination system is adopted to effectively avoid the risk of contamination of amplification products.
05.
Internal standard system
Introduce internal quality control (internal standards) to monitor the entire process of extraction and amplification, and improve the quality of detection.
06.
Professional
Calibrators can be traced back to digital PCR, ensuring accurate measurement values.
07.
Durability
The kit was repeatedly frozen and thawed 3, 5, 7 and 10 times, and the performance of the reagents was not affected.
08.
Instrument applicability
The performance is not affected on different models of devices such as ABI7500, ABlQuantStudio™5, Bio-RAD CFX96, SLAN-96P, SLAN-96S, and Roche Lightcycler480Ⅱ。
Related products 1 / Item Number: BP-QN28-100 rcAAV-2/N Detection Kit (PCR-Fluorescence Probe Method) 2 / Item Number: BP-QN36-100 Nucleic Acid Extraction or Purification Kit III (Magnetic Bead Method)
Scan the code for the reagent kit to consult
References
"Technical Guidelines for Pharmaceutical Research and Evaluation of Gene Therapy Products in Vivo (Trial)"
2. Technical Guidelines for Non-clinical Research and Evaluation of Gene Therapy Products
0756-8699969
Address: No. 88, Shuian 1st Road, Nanping Science and Technology Park, Xiangzhou District, Zhuhai City, Guangdong Province
Email: marketing@biori.com
Wechat official account |
Product consultation |
Join us |
Video Account |
