122nd General Meeting of the KCS

Type Oral Presentation
Area Oral Presentation of Young Analytical Chemists II
Room No. Room 321
Time FRI 09:15-:
Code ANAL2.O-2
Subject Simultaneous Determination of Highly Acidic Glycans in Biotherapeutics using a Combination of PGC-SPE and LC-MS/MS
Authors Youngsuk Seo, Hyun Joo An*
Graduate School of Analytical Science and Technology, Chungnam National University, Korea
Abstract Glycosylation of therapeutic glycoproteins affects the pharmacological functions including efficacy, safety, and biological activity of the drug. During the manufacturing process, various media-experimental conditions can lead to alterations in glycosylation. Therefore, overall glycan profiling, including both targeted and untargeted species, should be performed for drug QA/QC. In particular, acidic glycans including sialylated and phosphorylated glycans were directly associated with in vivo functions such as extended serum half-life, protein transfer, and immunogenicity. Despite the biological importance of acidic glycans, their analysis is still highly challenging due to the different physico-chemical properties. Here, we developed an efficient strategy providing high separation selectivity for simultaneous analysis of various acidic glycans using the combination of porous graphitized carbon-solid phase extraction (PGC-SPE) and high resolution LC/MS. For enhanced separation performance, diverse acidic glycans were pre-fractionated according to molecular size and polarity (acidity) using SPE with a PGC cartridge. Subsequent LC-MS and -MS/MS analyses enabled us to obtain glycan compositions, structures, and quantitative information of acidic glycans. We performed glycomic characterization of acidic glycans in a representative therapeutic enzyme, agalsidase-beta, to verify the method. Phosphorylated and sialylated glycans were successfully determined with high selectivity without any ion suppression/interference during LC/MS analysis. We have found high similarities to acid glycans between different batches of therapeutic enzymes. These results show that our method can be applied to assess the equivalence of biological drugs.
E-mail ysseo910@gmail.com