| Product Description |
Pembrolizumab, with the brand name Keytruda, is a humanized monoclonal
antibody of IgG4 isotype. It targets the programmed cell death protein 1 (PD-1)
receptor of lymphocytes, allowing the immune system to destroy cancer
cells. Pembrolizumab is an immunotherapy drug. It is used for the
treatment of patients with lung cancer, head and neck cancer, and stomach
cancer.
GenScript’s Pembrolizumab Pharmacokinetic ELISA Kit had been comprehensively
developed and validated for quantitative measurement of Pembrolizumab in
cynomolgus monkey serum and plasma, based on the ICH M10 and the FDA bioanalytical method validation
guidance for industry. Its precision, accuracy, dilutional linearity,
specificity, selectivity, stability, and hook effect were acceptable according
to the guidances[1-4]. The Pembrolizumab ELISA kit is a validated
tool for whole Pembrolizumab and its biosimilar quantification in biological
matrices for drug research and development. The
kit can be also used for the detection of single chain variable fragment (scFv)
of Pembrolizumab. |
| Linearity Range |
5 ng/mL-160 ng/mL |
| Precision |
Intra-assay: CV≤10%
Inter-assay: CV≤15% |
| Minimum required dilution (MRD) |
1:100 selected by cynomolgus monkey plasma |
| Specificity |
No cross-reactivity at 1600 ng/mL of Human IgG4 |
| Hook Effect |
Not observed at 15000 ng/mL of Pembrolizumab |
| Kit Contents |
| Component |
Quantity/Size |
Part No. |
| Capture Plate |
1 plate |
H1-80 |
| Standard Stock |
1 vial (50 μL) |
H1-10 |
| Sample Dilution Buffer |
1 bottle (60 mL) |
E1-90 |
| Biotin Anti-Pembrolizumab Antibody |
1 bottle (12 mL) |
H1-20 |
| Streptavidin-HRP |
1 bottle (12 mL) |
H1-30 |
| TMB Solution |
1 bottle (12 mL) |
A1-40 |
| 20× Wash Solution |
1 bottle (60 mL) |
A1-70 |
| Stop Solution |
1 bottle (6 mL) |
A1-50 |
| Plate Sealer |
2 pieces |
N/A |
|
| 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 |
Pembrolizumab Pharmacokinetic ELISA Kit is a classic bridging ELISA assay with a pair of
anti-idiotypic monoclonal capture and detection antibodies. When standards or
samples are added to the capture plate, the Pembrolizumab in the sample can be
captured on the plate coated with the Pembrolizumab capture antibody. Then the Biotin
Anti-Pembrolizumab Antibody is added to interact with the Pembrolizumab bound
on the plate. Streptavidin-HRP (Streptavidin-Horseradish Peroxidase conjugate) is
added to interact with the Biotin Anti-Pembrolizumab Antibody. After the washing
steps, TMB Solution is added, resulting in the formation of blue color. The
reaction is stopped by adding Stop Solution. Application of the Stop Solution
results in the color changing from blue to yellow. The intensity of the color can
be read at 450 nm and 630 nm by a microplate reader.
The quantity of Pembrolizumab
in the sample is precisely quantified against a Pembrolizumab standard curve. |
| Reference |
1, International Council for Harmonisation of TechnicalRequirements for Pharmaceuticals for Human Use. ICH Harmonised Guideline,Bioanalytical Method Validation M10 (2019).
2, US FDA. Bioanalytical Method Validation Guidance for Industry (2018).
3, European Medicines Agency. Guideline on Bioanalytical Method Validation(2011).
4, DeSilva B, Smith W, Weiner R et al. Recommendations for the bioanalyticalmethod validation of ligand-binding assays to support. |
Figure 1: Pembrolizumab ELISA kit standard curve.
A set of Pembrolizumab (PBZ) calibration standards from 160 ng/mL to 2.5 ng/mL was then diluted with Sample Dilution Buffer with a volume ratio of 1:100.
For
research use only. Not intended for human and animal therapeutic or diagnostic
use.
