A comparative study of monosaccharide composition analysis as a carbohydrate test for biopharmaceuticals

Harazono, Akira; Kobayashi, Tetsu; Kawasaki, Nana; Itoh, Seiga; Tada, Minoru; Hashii, Noritaka; Ishii, Akiko; Arato, Teruyo; Yanagihara, Shigehiro; Yagi, Yuki; Koga, Akihisa; Tsuda, Yuriko; Kimura, Mikiko; Sakita, Masashi; Kitamura, Satoshi; Yamaguchi, Hideto; Mimura, Hisashi; Murata, Yoshimi; Hamazume, Yasuki; Sato, Takayuki; Natsuka, Shunji; Kakehi, Kazuaki; Kinoshita, Mitsuhiro; Watanabe, Sakie; Yamaguchi, Teruhide

doi:10.1016/j.biologicals.2011.04.002

Cited by 60 publications

(35 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…pCO2, glucose, lactate and ammonia were measured by Bioprofile 400 (Nova Biomedical, Walthan, MA, USA). Galactose concentration was analyzed by high pH anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) method (Harazono et al 2011). Cell culture supernatants, after centrifugation, were aliquoted and kept frozen at -80°C for later analysis.…”

Section: Cell Count and Cell Metabolite Measurementmentioning

confidence: 99%

Galactose supplementation enhance sialylation of recombinant Fc-fusion protein in CHO cell: an insight into the role of galactosylation in sialylation

Liu

Wang

Fan

et al. 2015

World J Microbiol Biotechnol

View full text Add to dashboard Cite

Sialic acid levels of therapeutic glycoprotein play an important role in plasma half-life. An undesirable decrease of sialic acid content was observed when we increased Fc-fusion protein productivity fourfold in a GS-CHO cell line by bioprocess optimization. We investigated the potential mechanism for the sialic acid content reduction. We found that limited nucleotide sugar precursor and the extracellular sialidase were not responsible for the reduction of the sialic acid content after titer improvement. Oligosaccharide analysis revealed that the lack of protein galactosylation was the potential cause for the reduction of sialic acid content. Thus we validated this notion by evaluated galactose supplementation in 2 L bioreactors. Cell culture performance was not impacted by addition of up to 40 mM galactose except for the glucose consumption rate. Addition of 20 mM galactose to the bioreactor resulted in the increase of 44 % for total sialic acid content and 20.3 % for sialylated glycans. These data were further validated when the process was run on 200 L scaled bioreactor. These data together show that the galactosylation plays an apparent role in sialylation in our current system.

show abstract

Section: Cell Count and Cell Metabolite Measurementmentioning

confidence: 99%

Galactose supplementation enhance sialylation of recombinant Fc-fusion protein in CHO cell: an insight into the role of galactosylation in sialylation

Liu

Wang

Fan

et al. 2015

World J Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…Particularly, glycosylated biopharmaceuticals contain various glycoforms, and the heterogeneity of carbohydrate moiety is altered according to the manufacturing process. For this reason, glycan analysis is an essential part of protein characterization in development and quality control of biopharmaceuticals as well (Harazono et al, 2011). Therefore, the study of protein with post-translational modifications has increased explosively in recent proteomic research, and this phenomenon represents importance of glycans in this area (Wei and Li, 2009).…”

Section: Glycansmentioning

confidence: 99%

“…Though the advanced mass spectrometry plays a versatile role in current glycan analysis, due to the complexity of structure and the difficulty of isolation of glycans, there is no single method which can clearly and efficiently analyze all of them (Zaia, 2010). The glycan analysis is processed by a combination of several methods, such as monosaccharide composition analysis after the hydrolysis (Harazono et al, 2011), profiling of liberated glycans (Pabst and Altmann, 2011), and structural assignment (Zaia, 2010). A number of methods have been developed for glycomics using either chromatographic, electromigratory or mass spectrometric method as qualitative and quantitative methods.…”

Section: Glycansmentioning

confidence: 99%

Development of Metrology for Modern Biology

Park¹,

Choi²,

Jeong³

2012

Modern Metrology Concerns

View full text Add to dashboard Cite

“…Although there are previous literature reports that describe acid hydrolysis of glycoproteins into their constituent monosaccharide units with subsequent analysis using LC-MS [12][13][14][15][16][17] or capillary electrophoresis [18], each of these studies requires derivitization of monosaccharides for detection, likely incorporating measurement bias in proportion to the labeling inefficiency. The current industry standard for analysis of non-derivatized, hydrolyzed sugars -anion exchange chromatography with pulsed amperometric detection [19] (HPAEC-PAD) -is capable of chromatographic resolution of monosaccharides and can be a very sensitive detection technique, but is inherently incompatible with direct mass spectrometry detection, and therefore incompatible with isotope dilution techniques.…”

Section: Introductionmentioning

confidence: 98%

Separation of monosaccharides hydrolyzed from glycoproteins without the need for derivatization

2015

View full text Add to dashboard Cite

Chromatographic separation of monosaccharides hydrolyzed from glycoconjugates or complex, aggregate biomaterials, can be achieved by classic analytical methods without a need for derivatizing the monosaccharide subunits. A simple and sensitive method is presented for characterizing underivatized monosaccharides following hydrolysis from N- and O-linked glycoproteins using high-performance liquid chromatography separation with mass spectrometry detection (LC-MS). This method is adaptable for characterizing anything from purified glycoproteins to mixtures of glycoforms, for relative or absolute quantification applications, and even for the analysis of complex biomaterials. Use of an amide stationary phase with HILIC chromatography is demonstrated to retain the highly polar, underivatized monosaccharides and to resolve stereoisomers and potentially interfering contaminants. This work illustrates an original approach for characterization of N- and O-linked glycoprotein standards, mixtures, and for complex biological materials such as a total yeast extract.

show abstract

A comparative study of monosaccharide composition analysis as a carbohydrate test for biopharmaceuticals

Cited by 60 publications

References 30 publications

Galactose supplementation enhance sialylation of recombinant Fc-fusion protein in CHO cell: an insight into the role of galactosylation in sialylation

Galactose supplementation enhance sialylation of recombinant Fc-fusion protein in CHO cell: an insight into the role of galactosylation in sialylation

Development of Metrology for Modern Biology

Separation of monosaccharides hydrolyzed from glycoproteins without the need for derivatization

Contact Info

Product

Resources

About