|
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
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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 |
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