AAV Neutralizing Antibody Assay-AAV Service-CRO Service

In recent years, adeno-associated virus (AAV) has become a commonly used delivery vector tool for gene therapy drugs. However, in clinical treatment, AAV-specific antibodies can reduce the effectiveness of gene therapy to a certain extent, which greatly limits the promotion of AAV gene therapy. On the other hand, the potency of AAV antibodies may change over time in individuals, and the antibodies generated during treatment may also hinder patients from receiving the same gene therapy plan for secondary treatment, which will impose certain restrictions on the timeliness of gene therapy drugs. Therefore, it is particularly important to evaluate the presence and level of AAV immunity in patients through neutralizing antibody (NAb) assay during clinical treatment.

Figure 1. Mechanism of AAV neutralizing antibodies inhibiting vector transduction and transgene expression [1]

Applications:

● Preclinical testing: NAb testing can provide guidance for serotype selection, route of administration, and drug dosage of AAV gene therapy drugs.

● Clinical testing: Monitoring changes in NAbs can help evaluate whether the gene therapy drug has reached the effective dose level, efficacy, and duration during clinical treatment.

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1.AAV Neutralizing Antibody Test

We use various AAV serotypes to simulate the biological process of antibodies blocking the virus from entering cells. Through this process, we can detect whether the serum/antibody has neutralizing activity against the AAV virus and calculate the neutralizing potency of AAV antibodies in the serum/antibody.

● Cell ion: HEK 293, HeLa, etc.

● Test protocol:

After co-incubation with gradient-diluted serum/antibody to be tested, the AAV virus is used to infect cells. After 48 hours, the fluorescence intensity of each serum/antibody dilution level is measured using a full-wavelength multifunctional microplate reader. The neutralization effect of the serum/antibody to be tested is evaluated by comparing the viral infection status with that of the control without serum/antibody and analyzing the IC50 values.

● Customer sample requirements:

a. Sample type: serum, body fluid, etc., shipped on dry ice;

b. Sample volume: > 30 μl (can be customized for special requirements);

c. Sample requirements: sterile, and tested negative for hepatitis B, hepatitis C, HIV, syphilis, Epstein-Barr virus (EB), cytomegalovirus (CMV), etc.

Our advantages:

we use the Luciferase reporter gene system to simulate the biological response of antibodies blocking the virus from entering cells. This system provides a more intuitive and systematic way to reflect the neutralization process, and it has high sensitivity, specificity, throughput, and accuracy.

Neutralizing antibody test data presentation:

Figure 2. Diagram of neutralizing antibody tests for AAV2, AAV5, AAV8, and AAV9 serotypes

2. AAV Serotype Testing

AAV Serotype Testing
AAV1	AAV1/2	AAV2	AAV2-7m8	AAV oligo001
AAV5	AAV6	AAV6m	AAV6.2	AAV7
AAV8	AAV9	rAAV2-retro	AAV-PHP.eB	AAV-B10
AAV-PHP.S	AAV-PAN	AAV-DJ	AAV-ShH10	AAV-ShH10(Y445F)
AAV-Rh10	AAV-Anc80L65	AAV-BR1	AAV-BI30	AAV-SCH9
AAV MaCPNS1	AAV MaCPNS2	mac	AAV DSS
For more serotypes, please contact us.

Our advantages:

(1) High sensitivity; (2) High specificity; (3) High throughput; (4) High accuracy

3.AAV Production for Various Testings

Gene: Luciferase/EGFP/mCherry
AAV1	AAV1/2	AAV2	AAV2-7m8	AAV oligo001
AAV5	AAV6	AAV6m	AAV6.2	AAV7
AAV8	AAV9	rAAV2-retro	AAV-PHP.eB	AAV-B10
AAV-PHP.S	AAV-PAN	AAV-DJ	AAV-ShH10	AAV-ShH10(Y445F)
AAV-Rh10	AAV-Anc80L65	AAV-BR1	AAV-BI30	AAV-SCH9
AAV MaCPNS1	AAV MaCPNS2	mac	AAV DSS
For more information on the production of other serotypes of AAV viruses, please contact us.

Reference:

[1] Schulz M, Levy DI, Petropoulos CJ, Bashirians G, Winburn I, Mahn M, Somanathan S, Cheng SH, Byrne BJ. Binding and neutralizing anti-AAV antibodies: Detection and implications for rAAV-mediated gene therapy. Mol Ther. 2023 Mar 1;31(3):616-630.

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AAV Neutralizing Antibody Assay

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