Product Description |
VHH or nanobody, known for its high specificity, affinity, stability, and binding capacity, is commonly utilized in the affinity purification of antibodies, proteins, enzymes, etc. In gene therapy, VHH is the typical structure of affinity ligands used for the purification of AAV. It is crucial to detect residual VHH affinity ligands in the AAV product. GenScript has high-performance anti-VHH antibodies that can specifically bind to VHH. Based on this, GenScript has developed the high-sensitivity VHH Affinity Ligand ELISA Kit. The kit could detect the residual levels of various VHH affinity ligands and also be used for the quantitative analysis of VHH drugs. VHH Affinity Ligand ELISA Kit is an indispensable tool in gene therapy and VHH drug development. |
Assay Working Ranges |
Part No.
|
VHH Affinity Ligand standard
|
Typical assay working ranges (pg/mL)
|
Z03793
|
AAVX Affinity Ligand Standard
|
10-640
|
Z03794
|
AVB Affinity Ligand Standard
|
312.5-20,000
|
Z03795
|
AAV8 Affinity Ligand Standard
|
40-2,560
|
Z03796
|
KappaXP Affinity Ligand Standard
|
31.25-2,000
|
The kit includes the CaptureSelect™ (a trademark of Thermo Fisher Scientific) AAVX Affinity Ligand standard (a VHH affinity ligand), and the assay can be expanded by adding alternative standards, such as CaptureSelect™ AAV8 Affinity Ligand Standard, CaptureSelect™ KappaXP Affinity Ligand Standard, and Capto AVB Affinity Ligand Standard. The kit can also be used for a variety of other VHH affinity ligand residual assays or VHH drug quantification. Users can follow the protocol of the kit to evaluate the suitability of the test for other VHH.
|
Kit Contents |
Component |
Quantity/Size |
Part No. |
Capture Plate |
1 plate |
V1-80 |
Standard Stock |
1 vial (30 μL) |
V1-10 |
Biotin Anti-VHH Antibody |
1 vial (12 mL) |
V1-20 |
Streptavidin-HRP |
1 bottle (12 mL) |
V1-30 |
Sample Dilution Buffer |
1 bottle (60 mL) |
V1-60 |
TMB Solution |
1 bottle (12 mL) |
A1-40 |
20×Wash Solution |
1 bottle (60 mL) |
V1-70 |
Stop Solution |
1 bottle (6 mL) |
A1-50 |
Plate Sealer |
2 pieces |
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|
Storage |
The unopened kit is stable for at least 12 months from the date of manufacture at 2°C to 8°C, and the opened kit is stable for up to 1 month from the date of opening at 2°C to 8°C.
|
Assay Principle |
VHH Affinity Ligand ELISA Kit is based on the sandwich ELISA method. When standards or samples are added to the capture plate coated with the anti-VHH capture antibody, theVHHAffinity Ligand in the standards or samples can be captured on the plate. Then anti-VHH detection antibody conjugated with biotin is added to interact with the VHH Affinity Ligand bound on the plate. Streptavidin-Horseradish Peroxidase conjugate (Streptavidin-HRP) is added to interact with the biotin conjugated anti-VHH antibody (Biotin Anti-VHH Antibody). After washing steps, 3,3',5,5'-Tetramethylbenzidine solution (TMB Solution) is added to the plate and color development is stopped by addition of stop solution. Application of the Stop Solution results in the color changing from blue to yellow. The color intensity can be read at 450 nm by a microplate reader.The VHH standard curve can be used to accurately quantify the amount of VHH in the samples.
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Figure 1: IX. ASSAY PROCEDURE SUMMARY
Figure 2: CaptureSelect™ AAVX Affinity Ligand standard curve
Figure 3: AAVX Affinity Ligand can be accurately recovered in different AAV samples
For research use only. Not intended for human and animal therapeutic or diagnostic use.