Emerging mass spectrometry-based approaches to probe protein-receptor interactions: focus on overcoming physiological barriers

Abstract

Physiological barriers, such as the blood-brain barrier and intestinal epithelial barrier, remain significant obstacles towards wider utilization of biopharmaceutical products. Receptor-mediated transcytosis has long been viewed as an attractive means of crossing such barriers, but successful exploitation of this route requires better understanding of the interactions between the receptors and protein-based therapeutics. Detailed characterization of such processes at the molecular level is challenging due to the very large physical size and heterogeneity of these species, which makes use of many state-of-the art analytical techniques, such as high-resolution NMR and X-ray crystallography impractical. Mass spectrometry has emerged in the past decade as a powerful tool to study protein-receptor interactions, although its applications to investigate interaction of biopharmaceuticals with their physiological partners are still limited. We highlight the potential of this technique by considering several recent examples where it had been instrumental for understanding molecular mechanisms critical for receptor-mediated transcytosis of transferrin-based therapeutics.

Keywords: BBB; Blood–brain barrier; CNS; CSF; ESI; Fusion proteins; GHT; Growth hormone; HDX; Intestinal epithelial barrier; Lysozyme; Lz; LzT; MS; Protein aggregation; Protein therapeutics; Protein–drug conjugate; Receptor-mediated transcytosis; Tf; TfR; Transferrin; Transferrin receptor; central nervous system; cerebrospinal fluid; electrospray ionization; growth hormone-transferrin fusion protein; hydrogen/deuterium exchange; lysozyme; lysozyme–transferrin conjugate; mass spectrometry; transferrin; transferrin receptor.