원문정보
초록
영어
Quality control and assurance of glycan profiles of a recombinant glycoprotein from lot to lot is a critical issue in the pharmaceutical industry. To develop an easy and simple quantitative and qualitative glycan profile method based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), the modification with Girard’s reagent T (GT) was exploited. Since GT-derivatized quantification of oligosaccharides using MALDI-TOF MS is only possible with neutral glycans, sialylated glycans are not subjected to quantitative analysis with MALDI-TOF MS. To solve this problem, mild methyl esterification and subsequent GT derivatization were employed, which enabled us to perform rapid qualitative and quantitative analysis of sialylated and neutral N-linked oligosaccharides using MALDI-TOF MS. This modified method was utilized in the comparative quantification of N-glycans from the recombinant therapeutic glycoprotein expressed in two different kinds of chinese hamster ovary (CHO) cell line. The percentages of sialylated N-glycans to total were 22.5% and 5.2% in CHO-I cell and CHO-II, respectively, which resulted in significant difference in the biological activity of the recombinant glycoprotein.