膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| PAV005-C01 | Pre-coated AAV5 Capsid Protein Microplate | 1 plate |
| PAV005-C02 | Anti-AAV5 Antibody Standard | 20 μg |
| PAV005-C03 | Biotin-AAV5 Capsid Protein | 15 μg |
| PAV005-C04 | Streptavidin-HRP | 50 μL |
| PAV005-C05 | 10×Washing Buffer | 50 mL |
| PAV005-C06 | 2×Dilution Buffer | 50 mL |
| PAV005-C07 | Substrate Solution | 12 mL |
| PAV005-C08 | Stop Solution | 7 mL |
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）
Anti-AAV5 Antibody ELISA Kit is developed for the detection of anti-AAV5 antibodies in serum.It can be used for immunogenicity studies and enrollment screening.
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）
1. Unopened kit should be stored at 2℃-8℃ upon receiving.
2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理（Assay Principles）
This assay kit employs a sandwich-ELISA format, providing a rapid detection of Anti-AAV5 Antibody. The kit consists of Pre-coated AAV5 Capsid Protein Microplate, Anti-AAV5 Antibody Standard, Biotin-AAV5 Capsid Protein, Streptavidin-HRP and buffers.
Your experiment will include 6 simple steps:
a) All reagents were returned to room temperature(20℃-25℃) before use.
b) Add your sample to the plate, take the Anti-AAV5 Antibody as standard sample. The samples and standard sample are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-AAV5 Capsid Protein diluted by Dilution Buffer to the plate.
d) Wash the plate and add the diluted 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 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 Antibody bound.
典型数据-Typical Data Please refer to DS document for the assay protocol.
Detection of anti-AAV5 antibodies by sandwich-ELISA Assay.
Immobilized AAV5 Capsid Protein can bind Anti-AAV5 Antibody in 1:50 human serum, and then add Biotin-AAV5 Capsid Protein. Detection was performed using Streptavidin-HRP with sensitivity of 15.6 ng/mL.
The specific binding of PAV-A005 to Anti-AAV1/2/3/6/8/9 antibody was detected. The results showed that PAV-A005 only binds specifically to Anti-AAV5 antibody.
验证（Validation）
批内差异（Intra-Assay Statistics）
Three samples of known concentration in blank human serum were tested six times on one plate to assess intra-assay precision, Intra-Assay Precision CV≤15%.
Three samples of known concentration in blank mouse serum were tested six times on one plate to assess intra-assay precision, Intra-Assay Precision CV≤15%.
Three samples of known concentration in blank monkey serum were tested six times on one plate to assess intra-assay precision, Intra-Assay Precision CV≤15%.
批间差异（Inter-Assay Statistics）
Three samples of known concentration in blank human serum were tested in six separate assays to assess inter-assay precision, Inter-Assay Precision CV≤15%.
Three samples of known concentration in blank mouse serum were tested in six separate assays to assess inter-assay precision, Inter-Assay Precision CV≤15%.
Three samples of known concentration in blank monkey serum were tested in six separate assays to assess inter-assay precision, Inter-Assay Precision CV≤15%.
