A MALDI-TOF MS method for the simultaneous and quantitative analysis of neutral and sialylated glycans of CHO-expressed glycoproteins

被引：7

作者：

Tep, Samnang ^{[1
]}

Hincapie, Marina ^{[2
]}

Hancock, William S. ^{[2
]}

机构：

[1] Biogen Idec Inc, Cambridge Ctr 14, Cambridge, MA 02142 USA

[2] Northeastern Univ, Barnett Inst, Boston, MA 02115 USA

来源：

CARBOHYDRATE RESEARCH | 2012年 / 347卷 / 01期

关键词：

Isotopic label; Glycans; MALDI-TOF MS; N-ACETYLGLUCOSAMINE; BIOLOGICAL-ACTIVITY; RAPID METHOD; OLIGOSACCHARIDES; PROTEIN; QUANTIFICATION; GLYCOSYLATION; ANTIBODIES; CELLS;

D O I：

10.1016/j.carres.2011.10.005

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The development of a MALDI-TOF MS method for the quantitative analysis of the glycosylation of CHO-expressed biotherapeutic glycoproteins shall be presented. The method utilizes a well-established chemistry, reductive amination of glycans, to derivatize glycans with either a light analog (C-12(7) anthranilic acid) or a heavy analog ((13)C7 anthranilic acid) to allow for the direct comparison of the alternately-labeled glycans by MALDI-TOF MS. The method allows for the simultaneous analysis of neutral and sialylated glycans and displays a linear dynamic range over two orders of magnitude with sub-picomolar sensitivity. Additionally, because the glycans are derivatized with anthranilic acid, which is a very sensitive fluorophore, the glycans can be analyzed by chromatography with fluorescence detection. The need for this type of method is highlighted by the biotechnology/biopharmaceutical industry's continuous drive towards fully understanding process control. By providing this type of quantitative data, glycosylation changes of the expressed protein can be easily observed thereby helping to further advance the understanding of a major aspect of the biopharmaceutical process. (C) 2011 Elsevier Ltd. All rights reserved.

引用

页码：121 / 129

页数：9

共 34 条

[11] Recombinant antibody therapeutics: the impact of glycosylation on mechanisms of action [J].

Jefferis, Royston .

TRENDS IN PHARMACOLOGICAL SCIENCES, 2009, 30 (07) :356-362

[12] Getting the glycosylation right: Implications for the biotechnology industry [J].

Jenkins, N ;

Parekh, RB ;

James, DC .

NATURE BIOTECHNOLOGY, 1996, 14 (08) :975-981

[13] CARBOHYDRATES IN PROTEIN .3. PREPARATION AND SOME OF PROPERTIES OF A GLYCOPEPTIDE FROM HENS-EGG ALBUMIN [J].

JOHANSEN, PG ;

NEUBERGER, A ;

MARSHALL, RD .

BIOCHEMICAL JOURNAL, 1961, 78 (03) :518-&

[14] Selective clearance of glycoforms of a complex glycoprotein pharmaceutical caused by terminal N-acetylglucosamine is similar in humans and cynomolgus monkeys [J].

Jones, Andrew J. S. ;

Papac, Damon I. ;

Chin, Edward H. ;

Keck, Rodney ;

Baughman, Sharon A. ;

Lin, Yvonne S. ;

Kneer, Johannes ;

Battersby, John E. .

GLYCOBIOLOGY, 2007, 17 (05) :529-540

[15] Quantification in proteomics through stable isotope coding: A review [J].

Julka, S ;

Regnier, F .

JOURNAL OF PROTEOME RESEARCH, 2004, 3 (03) :350-363

[16] Differential protein expression profiling by iTRAO-2DLC-MS/MS of lung cancer cells undergoing epithelial-mesenchymal transition reveals a migratory/invasive phenotype [J].

Keshamouni, VG ;

Michailidis, G ;

Grasso, CS ;

Anthwal, S ;

Strahler, JR ;

Walker, A ;

Arenberg, DA ;

Reddy, RC ;

Akulapalli, S ;

Thannickal, VJ ;

Standiford, TJ ;

Andrews, PC ;

Omenn, GS .

JOURNAL OF PROTEOME RESEARCH, 2006, 5 (05) :1143-1154

[17] Rapid analysis of N-linked oligosaccharides in glycoproteins (ovalbumin, ribonuclease B and fetuin) by reversed-phase ultra-performance liquid chromatography with fluorescence detection and electrospray ionization time-of-flight mass spectrometry [J].

Kurihara, Takamasa ;

Min, Jun Zhe ;

Hirata, Asuka ;

Toyo'oka, Toshimasa ;

Inagaki, Shinsuke .

BIOMEDICAL CHROMATOGRAPHY, 2009, 23 (05) :516-523

[18] ROLE OF ANTENNARY STRUCTURE OF N-LINKED SUGAR CHAINS IN RENAL HANDLING OF RECOMBINANT-HUMAN-ERYTHROPOIETIN [J].

MISAIZU, T ;

MATSUKI, S ;

STRICKLAND, TW ;

TAKEUCHI, M ;

KOBATA, A ;

TAKASAKI, S .

BLOOD, 1995, 86 (11) :4097-4104

[19] IDAWG: Metabolic Incorporation of Stable Isotope Labels for Quantitative Glycomics of Cultured Cells [J].

Orlando, Ron ;

Lim, Jae-Min ;

Atwood, James A., III ;

Angel, Peggi M. ;

Fang, Meng ;

Aoki, Kazuhiro ;

Alvarez-Manilla, Gerardo ;

Moremen, Kelley W. ;

York, William S. ;

Tiemeyer, Michael ;

Pierce, Michael ;

Dalton, Stephen ;

Wells, Lance .

JOURNAL OF PROTEOME RESEARCH, 2009, 8 (08) :3816-3823

[20] Analysis of acidic oligosaccharides and glycopeptides by matrix assisted laser desorption ionization time-of-flight mass spectrometry [J].

Papac, DI ;

Wong, A ;

Jones, AJS .

ANALYTICAL CHEMISTRY, 1996, 68 (18) :3215-3223

← 1 2 3 4 →