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製品詳細
FAB
- Comprehensive validation - Validated with various antibody subtypes and antibody drugs.
- Simple and fast operation - No complicated washing steps, significantly reducing time.
- High batch consistency - Strict control over raw materials and finished product quality, ensuring a stable supply.
- Accurate and reliable results - High sensitivity with minimal matrix effects.
- High throughput capability - Support multiple product specifications, ideal for high-throughput screening.
- Fast completion - Results in just 1 hour.
Product Specifications
Assay TypeCompetition-TR-FRETReactivityHumanRegulatory StatusRUOAssay Time1 hrSample volume10 μLProduct Overview
The Human Fc gamma RIIIA / CD16a (F176) binding Kit (TR-FRET) is based on a homogeneous (no wash) competition TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer) to measure the interaction between human CD16a (F176) and antibody drug candidates or CD16a (F176) inhibitors. It is designed to facilitate the ADCC functional performance evaluation of antibody drug candidates, high-throughput screening of CD16a (F176) inhibitors within 0.5-1 hours. It can also be used as a universal detection tool to identify the ability of antibody drugs to bind to human CD16a (F176).
Storage
1. Unopened kit should be stored at 2℃-8℃ upon receiving.
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.
Materials Provided
IDComponentsSizeFRT06-C01Human Fc gamma RIIIA / CD16a (F176) Protein Europium-chelate100 tests/500 testsFRT06-C02FA labeled human IgG antibody100 tests/500 testsFRT06-C03Human IgG Standard400 μg/100 tests 2000 μg/500 testsFRT06-C04Sample Dilution Buffer10 mL/100 tests & 500 testsFRT06-C05Detection Buffer10 mL/100 tests & 500 testsAssay Principles
Human Fc gamma RIIIA / CD16a (F176) binding kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). Use the mixture of biotinylated human Fc gamma RIIIA / CD16a (F176) and Europium-chelate labeled streptavidin as the donor, FA labeled Human IgG1 antibody as the acceptor.
- In the absence of human Fc gamma RIIIA/CD16a (F176) binding components, the donor and acceptor are in close proximity due to the binding of human Fc gamma RIIIA/CD16a (F176) and FA-labeled Human IgG1 antibody. Upon excitation with a specific light source, the donor emits a 620 nm signal, which is absorbed by the acceptor, resulting in a 665 nm emission.
- In the presence of human Fc gamma RIIIA/CD16a (F176) binding components, they prevent the donor-acceptor interaction, inhibiting FRET from occurring.
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データ表示
Bioactivity-TR-FRET
Please refer to DS document for the assay protocol.
Inhibition Assay of interaction of Europium-chelate labeled human Fc gamma RIIIA / CD16a (F176) and FA labeled human lgG by Human IgG standard in a homogeneous (no wash) TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) competition assay, with a typical IC50 of 1.246 μM (QC tested).
The kit has been used to detect different subclasses of Human IgG (Human IgG1, Human IgG2, Human IgG3 and Human IgG4), which exhibit different IC50 results as expected. As shown in the figure, human CD16a (F176) binds to human IgG1, IgG2, IgG3 and IgG4 with low affinity, and IgG1 and IgG3 show the higher affinity than IgG2 and IgG4.
The kit has been used to detect different subclasses of mouse IgG (mouse IgG1, mouse IgG2a and mouse IgG2b), which exhibit different IC50 results as expected. As shown in the figure, human CD16a (F176) almost has no binding to mouse IgG1, mouse IgG2a and mouse IgG2b.
The kit has been used to detect four FDA approved antibody drugs with different affinities binding to human CD16a (F176). Bevacizumab and Efgartigimod alfa bind to human CD16a (F176) with the nanomolar affinity around 1000 nM. Toripalimab doesn’t bind to human CD16a (F176). The Fc of Eculizumab has been modified into the human IgG2 hinge region and human IgG4 CH2-CH3 region, so it doesn’t bind to human CD16a (F176).
Verify potential matrix effects by adding different levels of DEME, RPMI1640, FBS and HSA to the Sample Diluted buffer.
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最新の進展
医薬品開発の進捗- 英語名：
Low affinity immunoglobulin gamma Fc region receptor III-A
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- 市販の医薬品数：
0 詳細
- 治験医薬品数：
14 詳細
- 最新の開発ステージ：
Phase 2 Clinical
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