AAV Capsid ELISA Kit

Sample

Liquid

Species Reactivity

AAV

Detection Method

sELISA

Intended Use

The AAV Capsid ELISA Kit - Multiple Serotypes is a Sandwich ELISA Kit designed for quantitatively measuring the total capsid titer of multiple serotypes of adeno-associated virus (AAV) in testing samples during viral vector production. The kit is universally compatible with most serotypes of AAV, providing a ready-to-use and reproducible method for quantifying different serotypes of AAV.

It is recommended to use a positive control that matches the AAV capsids in the samples to generate an accurate standard curve for quantification. Please note that this kit provides standards for AAV2 only; standards for other serotypes are not included.

Contents of Kit

1. Capture Plate: 1 plate (8 wells x 12 strips); 12 strips are configured in plate; plate is sealed in a foil pouch with a desiccant.
2. AAV2 Control: 2 vials (Specified on product label) AAV2 Control is two vials of lyophilized powder containing empty AAV2 capsids. Reconstituted with deionized or distilled water before use. The number of empty AAV2 capsids in each batch of AAV2 Control is different, and this information is specified on the product label. The reconstituted AAV2 Control can be stored at 2°C to 8°C with the kit for up to 2 weeks. For
long term storage, please aliquot and store at -20°C or below. Avoid repeated freezing and thawing cycles
3. Detection Antibody: 1 bottle (12 mL)
4. Streptavidin-HRP: 1 bottle (12 mL
5. Sample Dilution Buffer: 1 bottle (60 mL)
6. 10× Lysis Buffer: 1 vial (1.5 mL)
7. 20× Wash Solution: 1 bottle (60 mL)
8. TMB Solution: 1 bottle (12 mL)
9. Stop Solution: 1 bottle (6 mL)
10. Plate Sealer: 2 pieces

Storage

The unopened kit is stable for at least 12 months from the date of manufacture at 2°C to 8°C, and the opened kit is stable for up to 1 month from the date of opening at 2°C to 8°C.

Performance Characteristics

Assay Working Ranges
The typical detection range of this kit for AAV2 is from 7.83E+07 to 7.0E+09 capsids/mL. The antibodies in this kit's raw material recognize AAV of different serotypes differently, so the typical assay working range of AAV varies when testing different serotypes, as shown above and Figure 3.
The antibody in this kit also recognizes AAVDJ and AAVrh.1, but this kit is not validated for the detection of these two AAVs. Although the remaining serotypes of AAV are not validated in this
kit, it does not necessarily mean that they cannot be detected. Users can follow the protocol
of this kit to evaluate the suitability of the test for other serotypes.

Typical assay working ranges for different AAV serotypes.
AAV2 AAV2 VLP 1.32E+08 - 7.00E+09
AAV1 AAV1 empty capsids 2.73E+07 - 1.75E+09
AAV5 AAV5 empty capsids 6.30E+07 - 4.00E+09
AAV6 AAV6 empty capsids 3.91E+07 - 2.50E+09
AAV8 AAV8 empty capsids 6.27E+07 - 4.00E+09
AAV9 AAV9 empty capsids 1.09E+08 - 7.00E+09


Figure 3: Overlay plot of standard curves for different AAV serotypes.

Precision

Intra-assay and inter-assay precision were measured in 3 pools of different concentrations, using 3 lots of kits.

Detection Limit

According to the evaluation method recommended in CLSI guideline EP17-A2, the LoB (Limit of Blank) of this kit is 3.50E+07 Capsids/mL.
According to the evaluation method recommended in CLSI guideline EP17-A2, the LoD (Limit of Detection) for AAV2 detection is 7.83E+07 Capsids/mL and the LoQ (Limit of Quantitation) for AAV2 detection is 1.16E+08 Capsids/mL.

General Description

AAV is now widely-used viral vectors for gene therapy. When using AAV as the viral vector in gene therapy, determination of the total capsid titer of the purified AAV particle product is an essential characterization procedure and a critical step in clinical applications.