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| Assay Type | Sandwich-ELISA |
| Analyte | AAV3 |
| Format | 96T |
| Regulatory Status | RUO |
| Sensitivity | <1.05E+08 capsids/mL |
| Standard Curve Range | 3.35E+09 capsids/mL-1.05E+08 capsids/mL |
| Assay Time | 2 hr 50 min |
| Suitable Sample Type | For the detection and quantitative determination of AAV3 capsid titration in AAV gene therapy product preparation processing. |
| Sample volume | 100 uL |
AAV3 Titration ELISA Kit was developed for the detection and quantitative determination of AAV3 capsid titration in AAV gene therapy product preparation processing.
It is for research use only.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
| ID | Components | Size |
| AAV03-C01 | Pre-coated Anti-AAV3 Antibody Microplate | 1 plate |
| AAV03-C02 | AAV3 Standard | 5.03E+10 capsids |
| AAV03-C03 | Biotin-Anti-AAV3 Antibody | 20 μg |
| AAV03-C04 | Streptavidin-HRP | 50 μL |
| AAV03-C05 | 10×Washing Buffer | 50 mL |
| AAV03-C06 | 2×Dilution Buffer | 50 mL |
| AAV03-C07 | Substrate Solution | 12 mL |
| AAV03-C08 | Stop Solution | 7 mL |
Your experiment will include 6 simple steps:
a) Bring all reagents to room temperature(20℃-25℃) before use.
b) Add your sample to the plate and take the AAV3 capsid as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-AAV3 Antibody diluted by Dilution Buffer to the plate.
d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.
e) Wash the plate and add TMB.
f) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated by the absorbance at 450 nm minus the absorbance at 630 nm to remove background disturbance before statistical analysis. The OD Value reflects the amount of bound protein.
For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.
To assess the linearity of the assay, samples spiked with high concentrations of AAV3 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.
Three samples of known concentration were tested in three separate assays to assess inter-assay precision.
Three AAV3 with different concentrations were tested to calculate the recovery rate.
The specific binding of AAV-A003 to AAV1/2/3/5/6/8/9/dj serotypes was detected. The results showed that AAV-A003 only binds specifically to AAV3.
The specific binding of AAV-A003 to native/denatured AAV3 was detected. The results showed that AAV-A003 only binds specifically to native AAV3. (Denatured AAV3: 95℃ for 10min)
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