Method for detecting infection titer of recombinant adeno-associated virus vector

By using a quantitative PCR method to directly detect recombinant adeno-associated virus vectors in infected cells, the operation process is simplified, and rapid, sensitive and accurate infection titer detection is achieved, solving the problems of cumbersome and time-consuming detection in existing technologies. It is suitable for 293T and HepG2.0 cells.

CN120829992APending Publication Date: 2025-10-24BEIJING SOLOBIO GENETECHNOLOGY CO LTD
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Patent Information

Application Number
CN202510465032.7
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Priority Date
2024-04-16
Filing Date
2025-04-15
Publication Date
2025-10-24

AI Technical Summary

Technical Problem

Existing methods for detecting the infectious titer of recombinant adeno-associated virus vectors are cumbersome, time-consuming, and have poor accuracy, and are unable to quickly and accurately reflect the amount of infectious active virus.

Method used

The recombinant adeno-associated virus vector sample is directly inoculated into infected cells. After culture, the cells are lysed and quantitative PCR is performed. The infection titer is calculated using specific primers and a standard curve, avoiding the helper virus and gradient dilution steps and simplifying the operation process.

Benefits of technology

It achieves rapid, sensitive and accurate detection of the infection titer of recombinant adeno-associated virus vectors, with high detection sensitivity, excellent accuracy and stability, and shortened detection time. It is suitable for 293T cells and HepG2.0 liver cells.

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Abstract

The invention discloses a method for detecting the infection titer of an AAV (adeno-associated virus) vector, which comprises the following steps: (1) adding a recombinant adeno-associated virus vector sample to be detected into cells, and culturing for a period of time; (2) cracking the infected cells to obtain a DNA sample; (3) performing quantitative PCR (qPCR) on the DNA sample to obtain the concentration of a target fragment in the recombinant adeno-associated virus vector sample; and (4) calculating the infection titer of the recombinant adeno-associated virus vector sample according to the concentration of the target fragment. The method provided by the invention is simpler and quicker, and can sensitively and accurately determine the infection titer of the infectious AAV vector in the biological product.
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Citation Information

Patent Citations

  • Method for detecting recombinant gland relevant viral vector infection titer

    CN101386891A

  • Recombinant adeno-associated viruses with enhanced liver tropism and uses thereof

    WO2022100748A1