FcRn binding kit

The FcRn binding kit is designed to monitor the binding of the IgG Fc region to the human FcRn receptor.

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The FcRn binding kit is designed to monitor the binding of the IgG Fc region to the human FcRn receptor.

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Overview

The neonatal Fc receptor is an Fc receptor found in the placenta. It displays the unique feature of facilitating IgG transport from the mother to the growing fetus. This receptor also has an important role in modulating IgG and albumin half-life in the bloodstream, influencing the pharmacokinetic parameters through an FcRn-mediated transcytosis process. The FcRn kit measures the binding between Fc-based molecules and the FcRn receptor.

Benefits

  • FAST ANTIBODY SELECTION
  • PHARMACOKINETIC PREDICTION
  • LOW SAMPLE CONSUMPTION

Assay principle

The FcRn receptor biochemical binding assay is a competitive assay involving the extracellular ectodomain of FcRn conjugated to terbium Cryptate, and human IgG1 labelled with d2. The unlabeled antibody or Fc fused drug competes with the d2 labelled IgG1 for binding to the receptor, resulting in a decrease in FRET signal.

Principle of the  HTRF FcRn binding assay

Assay protocol

The FcRn biochemical binding assay needs only 1 incubation step after reagent distribution. Terbium Cryptate pre-labelled FcRn ectodomain is dispensed, followed by the addition of the sample and IgG1 labelled with d2. After 2h of incubation at RT, the signal can be read.

Protocol of the  HTRF FcRn binding assay

FcRn binding kit species selectivity

IgG1 isotypes from human and mouse were serially diluted with the diluent buffer provided in the kit, starting from 1mg/mL. Human and Mouse IgG1 were then tested in a competition mode. The chart demonstrates that the human IgG1 antibody efficiently displaces the d2 labelled IgG1 antibody in a dose dependent manner, whereas the FRET signal remains constant when the mouse IgG1 concentration increases. These data indicate that the FcRn binding kit is human IgG specific and not compatible with other IgG species.

Validation of HTRF FcRn kit on human IgG1

Inhibitory effect of Rituximab and Pembrolizumab antibodies

The inhibitory effects of two FDA approved antibodies, anti-CD20 Rituximab and the anti-PD1 Pembrolizumab, were investigated with the FcRn binding kit. The anti-CD20 IgG1 Rituximab and the anti-PD1 IgG4 Pembrolizumab were serially diluted with the diluent buffer provided in the kit, starting from 1 mg/mL and tested in a competition dose response. The data presented in the graph show that both antibodies efficiently compete with the d2-labelled antibody in a dose-dependent manner, and display similar IC50. The results show that the kit is compatible with human IgG1 and IgG4 antibodies.

Validation of HTRF FcRn kit on Rituximab and Pembrolizumab 2 therapeutic antibodies

HTRF Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Product Insert FcRn Kit2 / 64FCRNPET-64FCRNPEG

64FCRNPET-64FCRNPEG - Product Insert

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