Grevys Algirdas, Nilsen Jeannette, Sand Kine M K, Daba Muluneh B, Øynebråten Inger, Bern Malin, McAdam Martin B, Foss Stian, Schlothauer Tilman, Michaelsen Terje E, Christianson Gregory J, Roopenian Derry C, Blumberg Richard S, Sandlie Inger, Andersen Jan Terje
Centre for Immune Regulation (CIR) and Department of Biosciences, University of Oslo, N-0316, Oslo, Norway.
CIR and Department of Immunology, Rikshospitalet, Oslo University Hospital and University of Oslo, PO Box 4950, N-0424, Oslo, Norway.
Nat Commun. 2018 Feb 12;9(1):621. doi: 10.1038/s41467-018-03061-x.
Albumin and IgG have remarkably long serum half-lives due to pH-dependent FcRn-mediated cellular recycling that rescues both ligands from intracellular degradation. Furthermore, increase in half-lives of IgG and albumin-based therapeutics has the potential to improve their efficacies, but there is a great need for robust methods for screening of relative FcRn-dependent recycling ability. Here, we report on a novel human endothelial cell-based recycling assay (HERA) that can be used for such pre-clinical screening. In HERA, rescue from degradation depends on FcRn, and engineered ligands are recycled in a manner that correlates with their half-lives in human FcRn transgenic mice. Thus, HERA is a novel cellular assay that can be used to predict how FcRn-binding proteins are rescued from intracellular degradation.
白蛋白和免疫球蛋白G（IgG）具有显著长的血清半衰期，这归因于pH依赖性的FcRn介导的细胞再循环，该过程可将两种配体从细胞内降解中拯救出来。此外，基于IgG和白蛋白的治疗药物半衰期的延长有可能提高其疗效，但迫切需要强大的方法来筛选相对的FcRn依赖性再循环能力。在此，我们报告了一种新型的基于人内皮细胞的再循环测定法（HERA），可用于此类临床前筛选。在HERA中，从降解中拯救出来取决于FcRn，并且工程化配体以与其在人FcRn转基因小鼠中的半衰期相关的方式进行再循环。因此，HERA是一种新型细胞测定法，可用于预测FcRn结合蛋白如何从细胞内降解中被拯救出来。
