| Product Description |
Trastuzumab, also known as Herceptin, is a
human epidermal growth factor receptor 2 (HER2) inhibitor for the treatment of
breast cancer. Trastuzumab is a recombinant humanized IgG1 kappa monoclonal
antibody. It binds to the extracellular domain of the HER2 protein on the on
the surface of HER2-positive tumor cells with high affinity. Trastuzumab can
suppress the cells growth and proliferation by blocking the ability of the
cancer cells to receive chemical signals.
Antibody drugs have the possibility to
generate anti-drug antibodies (ADAs), which may alter drug clearance and
neutralize target binding, causing reduction of drug efficacy. The
immunogenicity of antibody drugs may cause anaphylaxis, infusion reactions, and
immune complex disorders. Therefore, it is important to assess the presence and
impact of Trastuzumab ADAs on exposure, safety, and efficacy.
GenScript Trastuzumab Immunogenicity Kit (Bridging ELISA) is designed
for detection of anti-Trastuzumab antibody in serum and plasma samples. This
kit utilizes a validated bridging immunoassay method based on the FDA, EMA and NMPA
Immunogenicity Guidelines. Through rigorous validation studies, the kit has
been demonstrated high sensitivity, specificity, and is free from matrix
effects. It is an ideal tool for the analysis of ADA against Trastuzumab. |
| Sensitivity |
0.78 ng/mL
|
| Detection Range |
0.78-50 ng/mL
|
| Precision |
Intra-Assay: CV≤10%
Inter-Assay: CV≤15%
|
| Minimum required dilution (MRD) |
1:5, validated non-human primate plasma
|
| Hook Effect |
Not observed at 3,200 ng/mL of anti-Trastuzumab
antibodies
|
| Conveniency |
All reagents and buffers for the test are
provided and the test can be completed within 2 hours
|
| Kit Contents |
| Component |
Quantity/Size |
Part No. |
| Capture Plate |
1 plate |
R1-80 |
| Standard Stock |
1 vial (50 μL) |
R1-10 |
| Biotin Trastuzumab |
1 bottle (12 mL) |
R1-20 |
| Streptavidin-HRP |
1 bottle (12 mL) |
R1-30 |
| Sample Dilution Buffer |
1 bottle (60 mL) |
R1-60 |
| 20× Wash Solution |
1 bottle (60 mL) |
R1-70 |
| Stop Solution |
1 bottle (6 mL) |
A1-50 |
| TMB Solution |
1 bottle (12 mL) |
A1-40 |
| 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 |
The GenScript Trastuzumab
Immunogenicity ELISA Kit is a bridging immunoassay that utilizes a microplate coated with the Trastuzumab.
When standards or samples are added to the capture plate, anti-Trastuzumab
antibodies can be captured on the plate. The biotin-conjugated Trastuzumab is
then added to interact with the anti-Trastuzumab antibodies bound on the plate.
After a washing step, horseradish peroxidase conjugated streptavidin
(Streptavidin-HRP) is added and to react with the
3,3',5,5'-Tetramethylbenzidine solution (TMB Solution) to develop a blue
product in the solution. The reaction is stopped by adding stop solution, which
turns the color yellow and the absorbance can be read at 450 nm by a microplate
reader. The intensity of the reaction color is directly proportional to the
concentration of antibodies to Trastuzumab in samples. |
| Reference |
1.
Immunogenicity Testing of Therapeutic
Protein Products — Developing and Validating Assays for Anti-Drug Antibody
Detection.
2.
Immunogenicity of Trastuzumab in patients with
advanced tumors.
3.
Technical
Guideline on Immunogenicity of Therapeutic Agents, NMPA, 2021.03.
4.
Immunogenicity Testing of Therapeutic Protein
Products- Developing and Validating Assays for Anti-Drug Antibody Detection
Guidance for Industry, January 2019.
5.
Guideline on Immunogenicity assessment of
biotechnology-derived therapeutic proteins (EMEA/CHMP/BMWP/14327/2006 Rev 1),
1st Dec 2017. |
Figure 1: Trastuzumab Immunogenicity Kit (Bridging ELISA) standard curve.
Figure 2. MRD analysis of the Kit
For research use only.
Not intended for human and animal therapeutic or diagnostic use.
