A method of detecting aav antibody titers
By combining the CRISPR-Cas9 gene editing system with the AAV vector, the problems of low sensitivity and high cost in existing AAV antibody detection technologies have been solved, enabling accurate and convenient detection of AAV antibody titers.
CN116396983BActive Publication Date: 2026-05-26SUZHOU JINA XINGCHEN BIOTECHNOLOGY CO LTD
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SUZHOU JINA XINGCHEN BIOTECHNOLOGY CO LTD
- Filing Date
- 2022-10-10
- Publication Date
- 2026-05-26
AI Technical Summary
Technical Problem
Existing AAV antibody detection methods suffer from low sensitivity, high cost, complex operation, and difficulty in accurately detecting AAV antibody titers.
Method used
By combining the CRISPR-Cas9 gene editing system with the AAV vector, and by electroporating cells and detecting the expression efficiency of the reporter gene, a polynomial curve formula was established to accurately calculate the AAV antibody titer.
Benefits of technology
This technology enables the digital and quantitative detection of AAV antibody titers, simplifies the operation process, reduces detection costs, and improves the accuracy and reliability of the detection.
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Figure CN116396983B_ABST
Abstract
This invention provides a method for detecting AAV antibody titers, belonging to the field of biomedical technology, comprising: S1 preparing serum-free AAV vectors with different MOI values; preparing a mixture containing different volumes of test serum samples and AAV vectors with known MOI values for co-incubation; S2: adding plasmids containing the CRISPR-Cas9 gene editing system to electroporation buffer, performing electroporation on a known number of cells, adding the serum-free AAV vectors and mixture from step S1 to the electroporated cells respectively, and detecting the expression efficiency of the reporter gene after electroporation; S3: calculating the AAV antibody titer in the test serum sample based on the detection results. The main advantage of this invention is that it can display AAV antibody titers digitally and quantitatively, facilitating understanding and communication, and obtaining stable and reliable results using different cell lines and different AAV transduction doses. This method is easy to standardize and validate, saving time, effort, and detection costs compared to traditional methods, and directly reflects antibody neutralizing activity.
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