Jung Soo Kim (GRAST)

Hyun Joo An (GRAST)

Hyun Joo An (GRAST)

Jung Soo Kim (GRAST)

 In recent years, glycosylation of biopharmaceuticals has been recognized for its impact on their stability and efficacy, leading to extensive research to enhance their effectiveness through glycoengineering. However, there has been limited research on the glycosylation of receptor proteins that significantly impact the pharmacokinetics of biopharmaceuticals. Understanding the glycosylation and functions of these receptor proteins is crucial for accurate drug evaluation and the development of effective biopharmaceuticals. To address this gap, we selected protein X, which interacts with specific biopharmaceuticals and influence their pharmacokinetics. Our study shows that the function and localization of protein X at the cellular level vary with the presence or absence of its N-glycans. To analyze these N-glycans, we enzymatically released N-glycans from protein X expressed in human cells, followed by separation and enrichment using a PGC-SPE. The purified N-glycans were then analyzed using a PGC-nanoflow LC-MS/MS system and the data obtained were quantitatively compared according to glycan type. We found a unique glycan pattern in the N-glycans of protein X. LacdiNAc structures accounted for 47.93%, the structure with two fucoses attached to one HexNAc accounted for 17.17%, and fucosylated structures for 96.27%. These findings provide new insights into the role of glycosylation in protein X function and its potential impact on the pharmacokinetics of biopharmaceuticals.