Quantitative characterization of intact sialylated O-glycans with MALDI-MS for protein biotherapeutics
Tóm tắt
For validating O-glycosylation of protein biotherapeutics, we presented a quantitative O-glycomics method which is based on the neutralization of sialic acids, the specific release of O-glycans, and the introduction of permanent positive charge followed by quantitative MALDI-MS analysis. This method shows excellent technical reproducibility, linearity and sensitivity. In addition, it enables the quantification of intact O-glycans with minimal degradation or loss of sialic acids on these glycans compared to a conventional HPLC-based method. We then applied this method to quantitatively characterize O-glycans present on Etanercept. The analysis showed the relative abundances of mono- and di-sialylated core 1 O-glycans - were 79.3±0.8% and 17.3±1.4%, respectively. This glycomics technology could allow for the reliable quantitative analysis of intact O-glycans from glycoproteins and may contribute to validation of O-glycosylation protein biotherapeutics in the pharmaceutical industry.
Tài liệu tham khảo
S. Elliott, T. Lorenzini, S. Asher, K. Aoki, D. Brankow, L. Buck, L. Busse, D. Chang, J. Fuller, J. Grant, N. Hernday, M. Hokum, S. Hu, A. Knudten, N. Levin, R. Komorowski, F. Martin, R. Navarro, T. Osslund, G. Rogers, N. Rogers, G. Trail and J. Egrie, Nat. Biotechnol., 21, 414 (2003).
R. J. Sola and K. Griebenow, BioDrugs, 24, 9 (2010).
A. M. Sinclair and S. Elliott, J. Pharm. Sci., 94, 1626 (2005).
R. J. Sola and K. Griebenow, J. Pharm. Sci., 98, 1223 (2009).
P. Kang, Y. Mechref, Z. Kyselova, J. A. Goetz and M. V. Novotny, Anal. Chem., 79, 6064 (2007).
W. A. Tao and R. Aebersold, Curr. Opin. Biotechnol., 14, 110 (2003).
G. Alvarez-Manilla, N. L. Warren, T. Abney, J. Atwood, 3rd, P. Azadi, W. S. York, M. Pierce and R. Orlando, Glycobiology, 17, 677 (2007).
C. Wang, P. Zhang, W. Jin, L. Li, S. Qiang, Y. Zhang, L. Huang and Z. Wang, J. Proteomics, 150, 18 (2017).
M. Toyoda, H. Ito, Y. K. Matsuno, H. Narimatsu and A. Kameyama, Anal. Chem., 80, 5211 (2008).
G.C. Gil, B. Iliff, R. Cerny, W. H. Velander and K. E. Van Cott, Anal. Chem., 82, 6613 (2010).
H. J. Jeong, M. Adhya, H. M. Park, Y. G. Kim and B. G. Kim, Xenotransplantation, 20, 407 (2013).
C. Wang, Z. Wu, J. Yuan, B. Wang, P. Zhang, Y. Zhang, Z. Wang and L. Huang, J. Proteome. Res., 13, 372 (2014).
Y. Shinohara, J. Furukawa, K. Niikura, N. Miura and S. Nishimura, Anal. Chem., 76, 6989 (2004).
Y.W. Kim, C. Sung, S. Lee, K. J. Kim, Y. H. Yang, B. G. Kim, Y. K. Lee, H. W. Ryu and Y. G. Kim, Anal. Chem., 87, 858 (2015).
K. J. Kim, H. J. Kim, H. G. Park, C. H. Hwang, C. Sung, K. S. Jang, S. H. Park, B. G. Kim, Y. K. Lee, Y. H. Yang, J. H. Jeong and Y. G. Kim, Sci. Rep., 6, 24489 (2016).
K. J. Kim, Y. W. Kim, C. H. Hwang, H. G. Park, Y. H. Yang, M. Koo and Y. G. Kim, Biotechnol. Lett., 37, 2019 (2015).
L. Liu, S. Gomathinayagam, L. Hamuro, T. Prueksaritanont, W. Wang, T. A. Stadheim and S. R. Hamilton, Pharm. Res., 30, 803 (2013).
D. Pennica, V. T. Lam, R. F. Weber, W. J. Kohr, L. J. Basa, M. W. Spellman, A. Ashkenazi, S. J. Shire and D. V. Goeddel, Biochemistry, 32, 3131 (1993).
M. DiPaola, J. Li and E. Stephens, J. Bioanal. Biomed., 5, 5 (2013).
S. Houel, M. Hilliard, Y. Q. Yu, N. McLoughlin, S. M. Martin, P. M. Rudd, J. P. Williams and W. Chen, Anal. Chem., 86, 576 (2014).