earticle

영어

Glycan of glycoprotein drugs, which are over 70 % of recombinant DNA products, is a very important evaluation factor in product development or quality evaluation, because it was known as have important roles in both medicinal effects and stability. Especially, in case of biosimilar drugs, which are being recently actively developed, for proving similarity with their innovator, comparative analysis of glycan structure has come to the core as a fundamental evaluation factor. Therefore we desired to establish the useful scientific foundation for quality evaluation and approval evaluation of glycoprotein drugs, thereby establishing standardized glycan analysis methods for glycoprotein drugs using high-tech mass spectrometer in this study. To achieve this, we proposed glycan analysis methods and confirmed these methods, by testing human IgG as glycoprotein and on sale glycoprotein drugs(Infliximab and Trastuzumab). As a result, we established simplified sample preparation method in comparison time-consuming other researches and check method whether or not glycosylation using deglycosylation enzyme treatment and MALDI-TOF-MS. We also established glycosylation pattern and glycan sequence qualtitative analysis method using MALDI-TOF-MS, which has advantage of rapid analysis, and glycan relatively quantitative analysis method using LC-QTOF-MS, which has advantage of quantitative analysis. Established MALDI-TOF-MS method can use to check sample preparation properly and to analyze glycosylation pattern and glycan sequence qualitatively. For relatively quantitative analysis of glycan content and precisely qualitative analysis of glycosylation pattern and glycan sequence, LC-QTOF-MS is more effective than MALDI-TOF-MS. In addition, we proposed glycosylation site analysis method and confirmed the method using LC-QTOF-MS and glycan database. Through this study, we established total five SOPs(Standard Operating Procedures); ① Glycosylation ascertainment test by MALDI-MS, ② Glycan qualtitative analysis by MALDI-MS, ③ Glycan qualitative and quantitative analysis by LC-MS, ④ Glycosylation site analysis by LC-MS, ⑤ Sample preparation ascertainment test by MALDI-MS. These SOPs can be useful in characterizaion of biosimilars.