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Application: Screening inhibitors of FcRn binding to Fc (IgG1).
Background: Neonatal Fc receptor for IgG (FcRn) is a heterodimeric protein. FcRn consists of the Fc Gamma Receptor and Transporter encoded by the FCGRT gene, associated with beta-2-Microglobulin (B2M). FcRn binds to the Fc region of monomeric immunoglobulin G (IgG). It is expressed in over 25 tissue types, with high expression levels observed in the spleen and intestine. In the placenta, it transports IgGs from mother to fetus. FcRn contributes to an effective humoral immunity by protecting IgGs from degradation, recycling them and extending their half-life in circulation. In addition to IgGs, it regulates the homeostasis of serum albumin.The function of FcRn can be exploited by engineering therapeutic antibodies to increase their binding to FcRn, thereby improving their half-life and therapeutic efficacy. For example, an antibody cocktail that contains Fc mutations and an extended half-life (Evusheld) is used to treat COVID-19. The first-in-class drug, Enbrel, a TNF-alpha/Fc fuses Fc portions to a therapeutic protein to increase their half-life. There are now several other drugs in clinical using similar strategies.Conversely, FcRn is a potential therapeutic target for autoimmune diseases. Disrupting the FcRn/IgG interaction is expected to increase the overall clearance of IgGs, including disease-causing autoantibodies. Engineered Fc fragments or neutralizing IgGs that bind to FcRn with high affinity through their Fc region are currently undergoing clinical trial. The first FDA-approved drug targeting FcRn (efgartigimod) is now used to treat myasthenia gravis, an autoimmune neuromuscular disease caused by the presence of autoantibodies against acetylcholine receptor, providing proof-of-concept in favor of this strategy.
Contraindications: The DMSO concentration in the final reaction should be ≤1%.
Description: The Fc (IgG1): FcRn Inhibitor Screening Colorimetric Assay Kit is designed for the screening and profiling of neutralizing antibodies or inhibitors of the interaction between human Fc (IgG1) and human FcRn (Neonatal Fc receptor for IgG). This kit comes in a convenient 96-well or 384-well format, with purified Biotinylated-FcRn complex (Fc receptor amino acids 24-297 and B2M amino acids 21-119) and Fc (IgG1) (amino acids 100-330) proteins, Streptavidin-HRP, and assay buffers for 100 or 400 reactions. A 96-well plate is coated with Fc (IgG1) protein. After blocking, the plate is pre-incubated with an inhibitor or neutralizing antibody. Upon subsequent incubation with FcRn-biotin, the plate is treated with Streptavidin-HRP followed by addition of a colorimetric HRP substrate to produce color, which can be quenched and measured using a UV/Vis microplate reader. The signal is proportional to the binding of FcRn to Fc (IgG1).
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: This kit comes in a convenient 96-well format, with purified Biotinylated-FcRn and Fc (IgG1) proteins, Streptavidin-HRP, and assay buffers for 100 reactions.
Uniprot: P55899
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Dall'Acqua W.F., et al. 2002 J Immunol. 169(9): 5171-80
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