FcRn Reagents Platform Compatibility
Immunitrack’s FcRn reagents have been tested and validated by our customers and partners on some of the most popular high-throughput interaction assay platforms available. These include the AlphaScreen assay system (Perkin Elmer), the Octet® assay platform (ForteBio), and Biacore surface plasmon resonance-based biosensors (GE Healthcare). This broad platform compatibility makes it possible to use the FcRn reagents in most laboratories. See below for example data about how our FcRn reagents perform on these platforms.
Figure 1: Biotinylation of FcRn Reagents. The SDS-PAGE gel shift assays above show the biotinylation of FcRn heavy chains. Site-specific biotinylation at the C terminus ensures directed immobilisation and allows simultaneous binding of albumin and IgG. Immunitrack’s FcRn reagents are available to order in biotinylated and non-biotinylated formats.
Platform Compatibility – AlphaScreen (Perkin Elmer)
Figure 2: Titration of Humira®, which exhibits specificity for the human CD20 protein (green, Kd 32nM) and human serum albumin (HSA, orange, Kd 850) with biotinylated human FcRn using the Alpha Screen platform from Perkin Elmer. The Y-axis displays the assay signal expressed as a percentage of Bmax, while the X-axis shows the concentration of Humira and HSA in nanomolar (nm).
Platform Compatibility – Octet® (ForteBio)
Figure 3: IgG loading optimisation for FcRn assay on FAB2G biosensor. A) Raw data traces. In the loading step, FAB2G sensor is loaded with a 2-fold dilution series of IgG (ligand) starting at 25 µg/mL. A zero-IgG control is also run to assess non-specific binding of FcRn to the biosensor. In the analyte association step, FcRn is run with each IgG concentration at a single concentration of 1.6 uM (see application note).
Download an application note on the Octet Platform ▸
Platform Compatibility – Biacore (GE Healthcare)
Figure 4: Binding profiles for infliximab to human, macaque and mouse FcRn. Binding behaviour towards human and macaque FcRn is very similar. Infliximab exhibited greatest affinity for mouse FcRn. Binding specificities were in agreement with published data. See Biacore GE Healthcare application note for more details.
Download an application note on the Biacore Platform ▸
We realise that not all of our current and future customers are using the platforms mentioned above. If you are using Immunitrack’s FcRn reagents on a platform or setup not mentioned here and would like to share protocols, notes or feedback about their performance, please feel free to get in touch with us using the contact information at the bottom of the page. We are very grateful for all customer feedback!
Abdiche, Y. N.; Yeung, Y. A.; Chaparro-Riggers, J.; Barman, I.; Strop, P.; Chin, S. M.; Pham, A.; Bolton, G.; McDonough, D.; Lindquist, K.; Pons, J.; Rajpal, A., The neonatal Fc receptor (FcRn) binds independently to both sites of the IgG homodimer with identical affinity. MAbs 2015, 7 (2), 331-43.