膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | Sandwich-ELISA |
| Analyte | AAV2 |
| Format | 96T |
| Regulatory Status | RUO |
| Sensitivity | <3.13E+07 capsids/mL |
| Standard Curve Range | 1.00E+09 capsids/mL-3.13E+07 capsids/mL |
| Assay Time | 3 hr 20 min |
| Suitable Sample Type | For the detection and quantitative detection and quantitative determination of AAV2 capsid titration in AAV gene therapy product preparation processing. |
| Sample volume | 100 μL |
背景（Background）
Adeno-associated virus (AAV) has become one of the most important gene vectors in the field of gene therapy due to its long-term expression, low toxicity, low immunogenicity, and high tissue specificity.Most successful AAV gene therapies for preclinical and clinical studies are limited to natural serotypes, but the presence of neutralizing antibodies against AAV remains a significant barrier to systemic delivery.
应用说明（Application）
AAV2 Titration ELISA Kit was developed for the detection and quantitative determination of AAV2 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 |
| AAV02-C01 | Pre-coated Anti-AAV2 Antibody Microplate | 1 plate |
| AAV02-C02 | AAV2 Standard | 5.00E+10 capsids |
| AAV02-C03 | Biotin-Anti-AAV2 Antibody | 15 μg |
| AAV02-C04 | Streptavidin-HRP | 50 μL |
| AAV02-C05 | 10×Washing Buffer | 50 mL |
| AAV02-C06 | 2×Dilution Buffer | 50 mL |
| AAV02-C07 | Substrate Solution | 12 mL |
| AAV02-C08 | Stop Solution | 7 mL |
原理（Assay Principles）
This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of AAV2 capsid. The kit consists of Pre-coated Anti-AAV2 Antibody Microplate and AAV2 Standard,Biotin-Anti-AAV2 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 AAV2 capsid as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-AAV2 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 AAV2 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 AAV2 with different concentrations were tested to calculate the recovery rate.
专属性（Specificity）
The specific binding of AAV-A002 to AAV1/2/3/5/6/8/9/dj serotypes was detected. The results showed that AAV-A002 only binds specifically to AAV2.
The specific binding of AAV-A002 to native/denatured AAV2 was detected. The results showed that AAV-A002 only binds specifically to native AAV2. (Denatured AAV2: 95℃ for 5min)
背景（Background）：AAV2
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.
