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Your Position: Home > Kits > AAV5 > AAV-A005

AAV5 Titration ELISA Kit

For research use only.

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Product Details
Assay TypeSandwich-ELISA
AnalyteAAV5
Format96T
Regulatory StatusRUO
Sensitivity< 1.70E+07 capsids/mL
Standard Curve Range1.09E+9 capsids/mL-1.70E+07 capsids/mL
Assay Time3 hr 20 min
Suitable Sample TypeFor the detection and quantitative determination of AAV5 capsid titration in AAV gene therapy product preparation processing.
Sample volume100 uL
Materials Provided
IDComponentsSize
AAV05-C01Pre-coated Anti-AAV5 Antibody Microplate1 plate
AAV05-C02AAV5 Standard8.7E+10capsids
AAV05-C03Biotin-Anti-AAV5 Antibody20 μg
AAV05-C04Streptavidin-HRP50 μL
AAV05-C0510xWashing Buffer 50 mL
AAV05-C062xDilution Buffer50 mL
AAV05-C07Substrate Solution12 mL
AAV05-C08Stop Solution7 mL
  • Background
    AAV5 Titration ELISA Kit is based on ELISA sandwich method and designed to detect and quantitative determine of AAV5 capsid titration in AAV gene therapy product preparation processing. It contains AAV5 empty capsids and a pair of antibodies against AAV5. 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 AAV5 capsid titration.
  • Application

    AAV5 Titration ELISA Kit was developed for the detection and quantitative determination of AAV5 capsid titration in AAV gene therapy product preparation processing.

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • 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.

  • Assay Principles
    This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of AAV5 capsid. The kit consists of Pre-coated Anti-AAV5 Antibody Microplate and AAV5 Standard,Biotin-Anti-AAV5 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 AAV5 capsid as standard. The samples and standard are diluted by Dilution Buffer.

    c) Wash the plate and add the Biotin-Anti-AAV5 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.

Typical Data Please refer to DS document for The assay protocol.
 AAV5 TYPICAL DATA

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.

Validation
Intra-Assay Statistics

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV<10%.

 AAV5 INTRA-ASSAY STATISTICS
Inter-Assay Statistics

Three samples of known concentration were tested in three separate assays to assess inter-assay precision, Inter-Assay Precision CV<10%.

 AAV5 INTER-ASSAY STATISTICS
Recovery

Three AAV5 capsid with different concentrations were tested to calculate the recovery rate.

 AAV5 RECOVERY
Specificity

The specific binding of AAV-A005 to AAV1/2/3/8/9/dj serotypes was detected. The results showed that AAV-A005 only binds specifically to AAV5.

 AAV5 SPECIFICITY

The specific binding of AAV-A005 to native/denatured AAV5 was detected. The results showed that AAV-A005 only binds specifically to native AAV5. (Denatured AAV5: 95℃ for 10min)

 AAV5 SPECIFICITY
  • Background: AAV5
    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.
  • Clinical and Translational Updates

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