Analysis of glycoprotein-derived oligosaccharides in glycoproteins detected on two-dimensional gel by capillary electrophoresis using on-line concentration method

Kamoda, Satoru; Nakanishi, Yasuharu; Kinoshita, Mitsuhiro; Ishikawa, Rei; Kakehi, Kazuaki

doi:10.1016/j.chroma.2005.08.052

Cited by 30 publications

(25 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It seems that sialylation reduces the mobility differences between glycoforms probably due to an overproportional increase in the hydrodynamic radius by the attachment of charged sialic acids. The same effect was observed in CSE analysis of AGP glycans [38]. This lowered resolution is the reason, why a number of articles appeared, where neuraminidases were used to simplify the glycan signal patterns (e.g., [26]).…”

Section: Capillary Electrophoresissupporting

confidence: 56%

“…In addition, AA allows milder conditions for labeling [30] and the label is compatible with further techniques like LC and MALDI [42]. In some cases, pressure injection was used, but higher detection limits for APTS-labeled glycans upon electrokinetic injection were demonstrated [38]. In some reports, the question, whether size sieving takes place or not remained open [14,17].…”

Section: Capillary Electrophoresismentioning

confidence: 97%

See 1 more Smart Citation

Protein glycosylation analysis with capillary-based electromigrative separation techniques

Pattky

Hühn

2010

Bioanal Rev

View full text Add to dashboard Cite

An impressive complexity is associated with glycoproteins due to the microheterogeneity of glycosylation as posttranslational modification giving rise to a vast number of isoforms. The full characterization of glycoproteins is difficult to achieve, and a number of analytical methods have to be combined for a detailed understanding of glycosylation. In this review, we focus on capillary electromigrative separation techniques in the formats capillary electrophoresis, micellar electrokinetic chromatography, and capillary sieving electrophoresis. These separation techniques can be applied to all levels of glycosylation analysis including intact glycoproteins, glycopeptides, and released glycans. We here discuss the separation characteristics for each method and the information that they can provide for each level. Detection issues, especially laser-induced fluorescence detection and mass spectrometry are taken into account. In addition, tables provide an overview on the achievements made from the very beginning of glycosylation research by electromigrative separation techniques. From the literature presented here it is clear, that glycosylation analysis by electromigrative separation techniques is on the edge of transition of basic research and method development towards applications. First proof-ofprinciple studies for in-depth glycoprotein characterization and clinical diagnosis are described. However, this overview also shows that many basic aspects of separation have not yet been fully understood and more research is necessary to be able to fully use the capabilities of electromigrative separation techniques.

show abstract

Section: Capillary Electrophoresissupporting

confidence: 56%

Section: Capillary Electrophoresismentioning

confidence: 97%

Protein glycosylation analysis with capillary-based electromigrative separation techniques

Pattky

Hühn

2010

Bioanal Rev

View full text Add to dashboard Cite

show abstract

“…The gels were dehydrated, and the spots were excised and placed in microcentrifuge tubes. In-gel digestion was then conducted by the method of Komoda et al 13) Briefly, PNGase F (2 units/4 ml, Roche, Mannheim, Germany) and 10 mM phosphate buffer (pH 7.0, 100 ml) were added to the gel sample, and the mixture was incubated at 37 C overnight. The glycans released from the gel were labeled with 2-aminopryridine (PA) using BlotGlyco 14) (Sumitomo Bakelite, Tokyo) according to the manufacturer's instructions.…”

Section: )mentioning

confidence: 99%

The Multiplicity ofN-Glycan Structures of Bovine Milk 18 kDa Lactophorin (Milk GlyCAM-1)

Inagaki

Nakaya

Nohara

et al. 2010

Bioscience, Biotechnology, and Biochemistry

View full text Add to dashboard Cite

“…Labeled oligosaccharides can then be successfully analyzed by CE with a helium-cadmium (He-Cd) LIF detector (ex. 325 nm, em.405 nm) using a DB-1 capillary in 100 mM Tris-borate buffer (pH 8.3) containing 10% polyethylenglycol as the running buffer [43]. The 2-AA labeling method uses an inexpensive and easily commercially available reagent; thus, it is superior in applicability and long-term robustness.…”

Section: Profiling Of 2-aa-labeled Oligosaccharides By Ce-lifmentioning

confidence: 99%

Evaluation of glycosylation for quality assurance of antibody pharmaceuticals by capillary electrophoresis

Kamoda¹,

Kakehi

2008

Electrophoresis

Self Cite

View full text Add to dashboard Cite

The development of monoclonal antibody (mAb) pharmaceuticals has rapidly generated in this decade, and there are currently 23 FDA-approved mAb pharmaceuticals. Carbohydrate chains in mAb pharmaceuticals play important roles for the expression of their biological activities such as antibody-dependent cellular cytotoxicity, whereas the oligosaccharide profile is easily changed depending on the manufacturing conditions. Therefore, the evaluation of carbohydrate chains in detail is quite important for quality control in the development of mAb from early phases, but oligosaccharides in mAb have high heterogeneity and full characterization of oligosaccharide moiety in mAb has been a challenging target. Capillary electrophoresis (CE) is a powerful tool for the evaluation of glycan occupancy and oligosaccharide profile. This review focuses mainly on the application of CE to the analytical studies on glycosylation in mAb pharmaceuticals developed in our laboratory. The related techniques including the full-image microchip isoelectric focusing and the newly developed mass spectrometry technologies are also shown. The combination of the techniques described in this review will be a good guide for the evaluation of glycosylation for quality assurance of antibody pharmaceuticals by CE from the early stage of the development to the marketing stage.

show abstract

Analysis of glycoprotein-derived oligosaccharides in glycoproteins detected on two-dimensional gel by capillary electrophoresis using on-line concentration method

Cited by 30 publications

References 38 publications

Protein glycosylation analysis with capillary-based electromigrative separation techniques

Protein glycosylation analysis with capillary-based electromigrative separation techniques

The Multiplicity ofN-Glycan Structures of Bovine Milk 18 kDa Lactophorin (Milk GlyCAM-1)

Evaluation of glycosylation for quality assurance of antibody pharmaceuticals by capillary electrophoresis

Contact Info

Product

Resources

About