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组分（Materials Provided）
IDComponentsSizeAAV02H-C01Pre-coated Anti-AAV2 Antibody Microplate1 plateAAV02H-C02AAV2 Standard5.00E+10 capsidsAAV02H-C03HRP-Anti-AAV2 Antibody15 μgAAV02H-C0410×Washing Buffer50 mLAAV02H-C052×Dilution Buffer50 mLAAV02H-C06Substrate Solution12 mLAAV02H-C07Stop Solution7 mL产品概述（Product Overview）
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）
2-8℃
原理（Assay Principles）
Human Fc gamma RIIIA / CD16a (V176) binding kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). Use the mixture of biotinylated human Fc gamma RIIIA / CD16a (V176) and Europium-chelate labeled streptavidin as the donor, FA labeled Human IgG1 antibody as the acceptor.
- In the absence of human Fc gamma RIIIA/CD16a (V176) binding components, the donor and acceptor are in close proximity due to the binding of human Fc gamma RIIIA/CD16a (V176) and FA-labeled Human IgG1 antibody. Upon excitation with a specific light source, the donor emits a 620 nm signal, which is absorbed by the acceptor, resulting in a 665 nm emission.
- In the presence of human Fc gamma RIIIA/CD16a (V176) binding components, they disrupt the donor-acceptor interaction, preventing FRET from occurring.
Your experiment will include 5 simple steps:
a) Bring all reagents and samples to room temperature (20℃-25℃) before use.
b) Add your sample to the plate, take the AAV2 capsid as Control sample. The samples and Control sample are diluted by Dilution Buffer.
c) Add the HRP-Anti-AAV2 Antibody diluted by Dilution Buffer to the plate.
d) Wash the plate and add TMB.
e) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated as the absorbance at 450 nm minus the absorbance at 630 nm to remove background prior to statistical analysis. The OD Value reflects the amount of protein bound.
质量管理控制体系（QMS）
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数据展示
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.
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-A002H to AAV1/2/3/5/6/8/9/dj serotypes was detected. The results showed that AAV-A002H only binds specifically to AAV2.
The specific binding of AAV-A002H to native/denatured AAV2 was detected. The results showed that AAV-A002H only binds specifically to native AAV2. (Denatured AAV2: 95℃ for 5min)
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