New Product Launch | TR-FRET FcRn binding kit

Author: Senior Product Manager of UA Views：10

In the antibody drug development field, half-life is a key metric for evaluating the drug-likeness of antibody molecules, and the interaction between FcRn (neonatal Fc receptor) and IgG directly determines the circulatory half-life of antibodies in vivo. Screening and optimizing antibody molecules and rapidly assessing the binding activity between candidate antibodies and FcRn is an unavoidable experimental step in early antibody screening and Fc structure engineering.

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Recent Advances

New Product Launch丨TR-FRET FcRn binding kit

In the antibody drug development field, half-life is a key indicator for evaluating the druggability of antibody molecules, and the interaction between FcRn (neonatal Fc receptor) and IgG directly determines the circulating half-life of antibodies in vivo. Screening and optimizing antibody molecules and rapidly evaluating the binding activity of candidate antibodies to FcRn are unavoidable experimental steps in early antibody screening and Fc structure engineering.

Traditional FcRn binding assays involve cumbersome procedures, numerous incubation and washing steps, and low throughput, making it difficult to meet the primary screening needs of large numbers of candidate molecules. Is there a time-saving, labor-saving, wash-free detection solution suitable for high-throughput screening? UniOne® TRFRET Human FcRn Binding Kit (Cat. No.: UA086165), based on homogeneous time-resolved fluorescence TRFRET technology, provides researchers with an efficient tool for detecting human FcRn-IgG interactions.

I. Technical Principle

The kit uses a competitive TRFRET homogeneous detection system, eliminating all washing steps throughout the process. In the system, Eu-labeled anti-Tag1 antibody serves as the donor and recognizes Tag1-FcRn protein; Ac-labeled IgG serves as the acceptor. When the donor and acceptor are in spatial proximity, fluorescence resonance energy transfer occurs, generating a signal at 665 nm. When the test antibody drug is present, it competes with Ac-labeled IgG for binding to FcRn protein. The stronger the test antibody binds to FcRn, the more Ac-IgG is competitively dissociated, and the lower the 665 nm detection signal. The degree of signal decrease directly corresponds to the binding capacity of the candidate antibody to FcRn and can be used to calculate IC50 and quantify molecular binding activity.

Simply put: signal strength = competitive binding strength. Through dose-response curves, the FcRn binding characteristics of different antibody molecules can be intuitively compared.

Figure: Working principle of UniOne TR-FRET FcRn binding kit

II. Kit Highlights

1. Homogeneous wash-free operation: No washing steps, reducing sample loss and experimental errors caused by manual operations. The 384-well plate high-throughput format is suitable for primary screening of large batches of candidate antibodies.

2. Human FcRn system: Uses Human FcRn protein to simulate the real human binding environment, suitable for evaluating the binding capacity of human antibodies and engineered Fc variant molecules.

3. Complete supporting components: The kit contains Tag1-FcRn protein, IgG calibrator, Eu-labeled antibody, Ac-labeled IgG, and detection buffer, available in multiple sizes from 100T to 10000T to meet needs from small-scale to large-scale screening. All components are stored at -80°C; aliquoting is recommended to avoid repeated freeze-thaw cycles.

4. Mature data analysis solution: The instructions directly provide formulas for Ratio, net signal, and CV values. With a TRFRET plate reader, excitation at 320/340 nm and dual-wavelength signal reading at 620 nm and 665 nm enable rapid output of key experimental parameters such as IC50 and S/B, which can be directly used for experimental report preparation.

5. Wide application scenarios: Early molecular screening of antibody drugs, validation of Fc domain engineering effects, comparison of FcRn binding activity among different IgG subtypes, and in vitro prediction of candidate molecule half-life.

III. Experimental Workflow

The overall experimental operation logic is clear. Using 384-well shallow white plates as the detection carrier, the operation reference is as follows:

Figure: Operation workflow of UniOne TR-FRET FcRn binding kit

IV. Experimental Data

The TRFRET microplate reader collects dual-wavelength fluorescence, applies the kit's built-in formula to calculate the Ratio, plots a dose-response curve, and obtains the IC50 value to complete the quantitative evaluation of FcRn binding activity.

Figure: Validation data of UniOne TR-FRET FcRn binding kit

Related Product Recommendations

UniOne^®TR-FRET Human FcRn Binding Kit
Product Name	Cat. No.	Size
UniOne^®TR-FRET Human FcRn Binding Kit	UA086165	500 points

*Detection reagents are available in the following sizes: 100 points, 500 points, 2500 points, 10000 points.

This article is reviewed and published by the technical expert team of UA

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