膜杰作
Star Staining
产品描述（Product Details）
| 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 |
背景（Background）
AAV3 Titration ELISA Kit is based on ELISA sandwich method and designed to detect and quantitative determine of AAV3 capsid titration in AAV gene therapy product preparation processing. It contains AAV3 empty capsids and a pair of antibodies against AAV3. Results are obtained by four parameter logistic curve that were parallel to the standard curves obtained. The verification results indicate that this kit can be used for the quantitative determination of AAV3 capsid titration.
应用说明（Application）
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.
重构方法（Reconstitution）
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储（Storage）
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
组分（Materials Provided）
| 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 |
原理（Assay Principles）
This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of AAV3 capsid. The kit consists of Pre-coated Anti-AAV3 Antibody Microplate and AAV3 Standard,Biotin-Anti-AAV3 Antibody,Streptavidin-HRP and buffers.
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.
典型数据-Typical Data Please refer to DS document for the assay protocol.
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.
验证（Validation）
稀释线性（Dilution Linearity）
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.
批内差异（Intra-Assay Statistics）
Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.
批间差异（Inter-Assay Statistics）
Three samples of known concentration were tested in three separate assays to assess inter-assay precision.
回收率（Recovery）
Three AAV3 with different concentrations were tested to calculate the recovery rate.
专属性（Specificity）
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)
背景（Background）：AAV3
The adeno-associated virus (AAV) is a small (25 nm), non-enveloped virus of the parvoviridae family, including 12 different AAV serotypes. In the parvoviridae family it belongs to the genus dependoparvovirus, because it needs the presence of a helper virus for replication and assembly. The icosahedral AAV capsid composed of the capsid proteins VP1, VP2 and VP3 contains a linear, single-stranded DNA genome of 4.7 kb.
