| Product Description |
GenScript
AAVX Titer Capsid ELISA Kit - Multiple Serotypes (Cat. No. L01035) 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.
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.
|
| Assay Working Ranges |
| Serotype | Typical assay working ranges (Capsids/mL) |
|---|
| AAV2 | 1.32E+08 - 7.00E+09 | | AAV1 | 2.73E+07 - 1.75E+09 | | AAV5 | 6.30E+07 - 4.00E+09 | | AAV6 | 3.91E+07 - 2.50E+09 | | AAV8 | 6.27E+07 - 4.00E+09 | | AAV9 | 1.09E+08 - 7.00E+09 |
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.
|
| Kit Contents |
| Component |
Quantity/Size |
Part No. |
| Capture Plate |
1 plate |
W1-80 |
| AAV2 Control |
2 vials (Specified on product label) |
W1-10 |
| Detection Antibody |
1 bottle (12 mL) |
W1-20 |
| Streptavidin-HRP |
1 bottle (12 mL) |
W1-30 |
| Sample Dilution Buffer |
1 bottle (60 mL) |
W1-60 |
| 10× Lysis Buffer |
1 vial (1.5 mL) |
W1-61 |
| 20× Wash Solution |
1 bottle (60 mL) |
W1-70 |
| TMB Solution |
1 bottle (12 mL) |
A1-40 |
| Stop Solution |
1 bottle (6 mL) |
A1-50 |
| 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. |
| Assay Principle |
AAVX
Titer Capsid ELISA Kit - Multiple Serotypes is based on the sandwich ELISA method.
When standards or samples are added to the capture plate coated with the
capture antibody that recognizes multiple serotypes, the AAV capsids
in the samples can be captured on the plate The detection antibody that recognizes
multiple serotypes and is conjugated with biotin, is then added to interact
with the AAV capsids bound on the plate. Streptavidin-Horseradish Peroxidase conjugate
is added to interact with the biotin conjugated detection antibody. After
washing steps, TMB solution is added to the plate and color development is
stopped by addition of stop solution. Application of the Stop Solution results
in the color changing from blue to yellow. The color intensity can be read at
450 nm by a microplate reader. The AAV standard curve can be used to accurately
quantify the amount of AAV capsids in the samples. |
| Reference |
1.
CLSI document EP17-A2 (Evaluation of Detection
Capability for Clinical Laboratory Measurement Procedures, 2nd Edition). |
A typical example of AAV2 standard curve using a four-parameter logistic curve. Each standard was analyzed in duplicate. The R-Square of this curve is 0.99977.
Overlay plot of standard curves for different AAV serotypes.
For research use only.
Not intended for human and animal therapeutic or diagnostic use.
