Expression, purification, and characterization of human mannose-6-phosphate receptor – Extra cellular domain from a stable cell line utilizing a small molecule biomimetic of the mannose-6-phosphate moiety

In this study, a novel human M6PR (hM6PR)-overexpressing cell line originally established for hM6PR cellular uptake assay was utilized for production of hM6PR-ECD, and a novel small molecule biomimetic (aminophenyl-M6P) affinity resin was developed for the purification of M6PR-ECD. The affinity-purified hM6PR-ECD was monomeric, contained 14 intact MRH domains (1–14) and a partial MRH domain 15, and was successfully employed in ELISA-based and surface plasmon resonance-based binding assays to demonstrate its ligand-binding functionality, making it suitable for the evaluation of biotherapeutics that utilize M6PR for cellular internalization.

Source: Protein Expression and Purification - February 4, 2020 Category: Biochemistry Source Type: research