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优势与特点（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.
产品参数（Product Specifications）
Assay TypeCompetition-TR-FRETReactivityHumanRegulatory StatusRUOAssay Time1 hrSample volume10 μL产品概述（Product Overview）
Human Fc gamma RI / CD64 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 CD64 and antibody drug candidates or CD64 inhibitors. It is designed to facilitate the ADCC and ADCP functional performance evaluation of antibody drug candidates, high-throughput screening of CD64 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 CD64.
存储（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.
组分（Materials Provided）
IDComponentsSizeFRT03-C01Human Fc gamma RI / CD64 Protein Europium-chelate100 tests/500 testsFRT03-C02FA labeled human IgG antibody100 tests/500 testsFRT03-C03Human IgG standard100 μg/100 tests 500 μg/500 testsDB-04TR-FRET Sample Dilution Buffer, pH7.450 mL/100 tests & 500 testsDB-05TR-FRET Detection Buffer, pH7.450 mL/100 tests & 500 tests原理（Assay Principles）
This Human Fc gamma RI / CD64 binding kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). This assay uses the mixture of biotinylated human Fc gamma RI / CD64 and Europium-chelate labeled streptavidin as the Donor and FA labeled human IgG antibody as the Acceptor.
— When the sample does not contain human Fc gamma RI / CD64 binding components, the Donor and Acceptor are in close proximity because of the binding of human Fc gamma RI / CD64 and human IgG antibody. Upon Donor excitation with light of a specific wavelength (337nm), in addition to Donor emission (620nm), non-radiative transfer of energy occurs between Donor and Acceptor, resulting in Acceptor emission (665nm).
— When the sample contains human Fc gamma RI / CD64 binding components, the components inhibit the binding between the Donor and Acceptor and thereby prevent FRET from occurring.
质量管理控制体系（QMS）
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数据展示
活性（Bioactivity）-TR-FRET
Please refer to DS document for the assay protocol.
In this TR-FRET assay, Human Fc gamma RI / CD64 Protein Europium-chelate is used as the Donor and FA labeled human IgG antibody is used as the Acceptor. An inhibition Assay was performed to evaluate the interaction between Human Fc gamma RI / CD64 Protein Europium-chelate and FA labeled human IgG antibody using Human IgG standard resulting in a typical IC50 of 5.903 nM (QC tested).
The kit can be used to detect different subclasses of Human IgG Whole and Fc fragment proteins (Human IgG1, Human IgG2, Human IgG3 and Human IgG4). As shown in the figure, human CD64 is a high affinity receptor that binds to human IgG1, IgG3 and IgG4, while it has not been shown to bind to human IgG2.
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 shown in the figure, human CD64 binds to mouse IgG2a with a high affinity, but not to mouse IgG1 and mouse IgG2b.
This kit was used to evaluate four FDA-approved antibody drugs with different human CD64 affinities. Bevacizumab, Toripalimab, and Efgartigimod alfa bind to human CD64 with high affinity. The Fc region of Eculizumab has been modified into the human IgG2 hinge region and the human IgG4 CH2-CH3 region, so it doesn’t bind to human CD64.
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