| Products/Services Used | Details | Operation |
|---|---|---|
| Recombinant Proteins | We coated Streptavidin MSD plates with a biotinylated anti- VHH antibody (Genscript A01995) at 2 μg/ml in PBS for 1 hour at RT with 1000 rpm, washed 3X with PBT, incubated with blocking buffer for 1 hour at RT, incubated with VHH-mFc containing media/mucus (at different dilutions) for 1 hour at RT with 1000 rpm, washed 3X with PBT, incubated with ruthenylated-anti-human-Fc antibody (Clone R10Z8E9, labelled in-house) at 2 ug/ml in PBS for 1 hour at RT with 1000 rpm, washed 3X with PBT and read plates in 40 μL reading buffer using the MSD imager.... The primary antibody mix contained mouse anti-pIgR antibody (R&D Systems, MAB27171) and biotinylated anti-VHH antibody (Genscript A01995) both at 5 μg/ml. | Get A Quote |
Mucosal immunity is dominated by secretory IgA and IgM, although these are less favorable compared to IgG molecules for therapeutic development. Polymeric IgA and IgM are actively transported across the epithelial barrier via engagement of the polymeric Ig receptor (pIgR), but IgG molecules lack a lumen-targeted active transport mechanism, resulting in poor biodistribution of IgG therapeutics in mucosal tissues. In this work, we describe the discovery and characterization of single-domain antibodies (VHH) that engage pIgR and undergo transepithelial transport across the mucosal epithelium. The anti-pIgR VHH panel displayed a broad range of biophysical characteristics, epitope diversity, IgA competition profiles... More
