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| Assay Type | Sandwich-ELISA |
| Analyte | Protein A |
| Format | 96T(8×12 strips) |
| Regulatory Status | RUO |
| Sensitivity | < 20pg/mL |
| Standard Curve Range | 25 pg/mL-1600 pg/mL |
| Assay Time | 2 hr |
| Suitable Sample Type | For the quantitative determination of recombinant Protein A, alkaline-resistant Protein A and MaXtar® ARPA ligand Protein A. |
| Sample volume | 50 μL |
Find the expiration date on the outside packaging and do not use reagents past their expiration date.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
| ID | Components | Size |
| RES024-C01 | Pre-Coated Anti-Protein A Antibody Microplate | 1 plate(8×12 strips) |
| RES024-C02A | Alkali-Tolerant Recombinant Protein A Standard (1μg/mL) | 100 μL |
| RES024-C02B | MaXtar® ARPA ligand Protein A Standard (Bio-Link Co.) (1μg/mL) | 100 μL |
| RES024-C03 | Recombinant Protein A Standard (1μg/mL) | 100 μL |
| RES024-C04 | Biotin-Anti-Protein A Antibody | 700 μL |
| RES024-C05 | Streptavidin-HRP | 300 μL |
| RES024-C06 | Dilution Buffer | 100 mL |
| RES024-C07 | 20×Washing Buffer | 30 mL |
| RES024-C08 | Substrate Solution | 12 mL |
| RES024-C09 | Stop Solution | 6 mL |
Detection of Recombinant Protein A by sandwich-ELISA Assay.
Immobilized Anti-Protein A Antibody can bind Recombinant Protein A. Detection was performed using Biotin-Anti-Protein A Antibody with sensitivity of 25 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.
Detection of Alkali-Tolerant Recombinant Protein A by sandwich-ELISA Assay.
Immobilized Anti-Protein A Antibody can bind Alkali-Tolerant Recombinant Protein A. Detection was performed using Biotin-Anti-Protein A Antibody with sensitivity of 25 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.
Detection of MaXtar® ARPA ligand Protein A (Bio-Link Co.) by sandwich-ELISA Assay.
Immobilized Anti-Protein A Antibody can bind MaXtar® ARPA ligand Protein A (Bio-Link Co.). Detection was performed using Biotin-Anti-Protein A Antibody with sensitivity of 25 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, Intra-Assay Precision CV＜10%.
Three samples of known concentration were tested in ten separate assays to assess inter-assay precision, Inter-Assay Precision CV<20%.
The following recovery tests were conducted by adding different concentrations of Protein A to Human IgG1（Bevacizumab）, and the final concentration of Human IgG1（Bevacizumab）is 10mg/mL to calculate the recovery rate. For Recombinant Protein A, Alkali-Tolerant Recombinant Protein A and MaXtar® ARPA ligand Protein A (Bio-Link Co.), the recovery ranges are 98-118%, 86-109% and 79-106%, respectively.
The following recovery tests were conducted by adding different concentrations of Protein A to Human IgG4（Toripalimab）, and the final concentration of Human IgG4（Toripalimab）is 10mg/mL to calculate the recovery rate. For Recombinant Protein A, Alkali-Tolerant Recombinant Protein A and MaXtar® ARPA ligand Protein A (Bio-Link Co.), the recovery ranges are 97-106%, 93-104% and 86-105%, respectively.
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) and host cell DNA (HCD) were added to human IgG1 (Bevacizumab) and human IgG4 (Toripalimab), respectively, which were higher than the usual quality standard limit. Then three samples of known concentration of Protein A were added, respectively, and the ratio of Protein A recovery in the Protein A added samples without HCP and HCD was added as the specificity verification index. The calculation formula was as follows: (S3-S1) / (S2-S1) × 100%, the experimental design is as follows.
The specificity results are as follows.
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