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| Assay Type | Sandwich-ELISA |
| Analyte | VH3 binding Protein A |
| Format | 96T |
| Regulatory Status | RUO |
| Sensitivity | ＜50pg/mL |
| Standard Curve Range | 50 pg/mL-8100 pg/mL |
| Assay Time | 2 hr |
| Suitable Sample Type | For the quantitative determination of recombinant VH3 binding protein A |
| Sample volume | 50ul |
The kit is developed for the detection of natural or structurally conserved recombinant forms of VH3 and a recombinant form of VH3 with very significant structural differences from natural VH3 such as MabSelect VH3 in Bioprocess manufacturing applications. It is used as a tool to aid in optimal purification process development and in routine quality control of in-process streams as well as final product .
It is for research use only.
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.
| ID | Components | Size |
| RES059-C01 | Pre-Coated Anti-VH3 Antibody Microplate | 1 plate |
| RES059-C02 | Recombinant VH3 binding Protein A Standard (1μg/mL) | 100uL |
| RES059-C03 | Biotin-Anti-VH3 Antibody | 1.5mL |
| RES059-C04 | Streptavidin-HRP | 10ug |
| RES059-C05 | 10×Sample Dilution Buffer | 15mL |
| RES059-C06 | Denaturation Buffer | 15mL |
| RES059-C07 | 20×Washing Buffer | 30mL |
| RES059-C08 | Antibody Dilution Buffer | 15mL |
| RES059-C09 | Streptavidin-HRP Dilution Buffer | 15mL |
| RES059-C10 | Substrate Solution | 12mL |
| RES059-C11 | Stop Solution | 8mL |
Detection of Recombinant VH3 binding Protein A by sandwich-ELISA Assay.
Immobilized Anti-VH3 binding Protein A Antibody can bind Recombinant VH3 binding Protein A. Detection was performed using Biotin-Anti-VH3 Antibody with sensitivity of 50 pg/mL (QC tested). 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.
Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.
Three samples of known concentration were tested in three separate assays to assess inter-assay precision.
Add different concentrations of VH3 binding Protein A (0.2ng/mL、1ng/mL、10ng/mL) to different concentrations of Human IgG4 (Toripalimab) (20mg/mL、10mg/mL、5mg/mL), then dilute the antibodies to a reasonable range, then test and calculated the concentration of VH3 binding protein A to give the recovery rate.
We have conducted interference effect test about frequently-used buffers, they have excellent buffer compatibility. For specific buffers, it is recommended that you verify recovery to determine the minimum dilution ratio.
Host cell protein (HCP 500 ng/mL) and host cell DNA (HCD 0.5 ng/mL) of HEK293, E.coli or CHO systems were added to human IgG1 (Bevacizumab, 1mg/mL) and human IgG4 (Toripalimab, 1mg/mL), respectively, which were higher than the usual quality standard limit. Then high, medium, and low concentrations of VH3 binding Protein A were added, respectively, and the ratio of VH3 binding Protein A recovery in the VH3 binding Protein A added samples without HCP and HCD was added as the specificity verification index. verification index.
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