Plug-plug kinetic capillary electrophoresis for in-capillary exoglycosidase digestion as a profiling tool for the analysis of glycoprotein glycans

Yamagami, Maki; Matsui, Yoshitaka; Hayakawa, Tetsuo; Yamamoto, Sachio; Kinoshita, Mitsuhiro; Suzuki, Shigeo

doi:10.1016/j.chroma.2017.03.019

Cited by 20 publications

(17 citation statements)

References 35 publications

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“…Hence, ACE possesses a great potential for investigation of the (bio)molecular noncovalent interactions under mild conditions and for estimation of binding (stability, association, formation) or dissociation constants of the formed complexes as demonstrated in several recent reviews and research articles . ACE involves several modes based either on the separation of the interacting species and determination of their equilibrium concentration, such as in the Hummel–Dreyer method , the vacancy peak method , frontal analysis , continuous frontal analysis , and kinetic CE , or on the detection of a specific physicochemical property of the complexed ligand or the binding partner, mostly the changes of effective mobility in the mobility shift assay or changes of migration times in the partial‐filling ACE . All these methods utilize the differences in the migration velocities of the interacting species.…”

Section: Separations By the Particular Ce And Cec Methodsmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Kašička

2017

Electrophoresis

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The review brings a comprehensive overview of recent developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) to analysis, microscale isolation, purification, and physicochemical and biochemical characterization of peptides in the years 2015, 2016, and ca. up to the middle of 2017. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis (sample preseparation, preconcentration and derivatization, adsorption suppression and EOF control, and detection) are described. New developments in particular CE and CEC methods are presented and several types of their applications to peptide analysis are reported: qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence and chiral analysis, and peptide mapping of proteins. Some micropreparative peptide separations are shown and capabilities of CE and CEC methods to provide important physicochemical characteristics of peptides are demonstrated.

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Section: Separations By the Particular Ce And Cec Methodsmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Kašička

2017

Electrophoresis

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“…Alternatively, on-line enzymatic reactions require nanoliter volumes of substrates and enzymes, and incubation times as low as 40 s have been reported [26]. Electroosmotic flow is typically suppressed through neutral coatings, such as poly(dimethylsiloxane) [98] or phospholipid [26,50], and background electrolytes with a higher viscosity than water. These viscosities were achieved through the use of hydroxypropylcellulose [98] or a phospholipid nanogel [26,50].…”

Section: Emerging Techniques and Future Directionsmentioning

confidence: 99%

“…Electroosmotic flow is typically suppressed through neutral coatings, such as poly(dimethylsiloxane) [98] or phospholipid [26,50], and background electrolytes with a higher viscosity than water. These viscosities were achieved through the use of hydroxypropylcellulose [98] or a phospholipid nanogel [26,50]. After optimization of several on-line reaction parameters, such as the length of the enzyme plug, enzyme concentration, and incubation time, the on-line sequencing of the terminal residues of complex N-glycans was achieved [26,50,98].…”

Section: Emerging Techniques and Future Directionsmentioning

confidence: 99%

“…These viscosities were achieved through the use of hydroxypropylcellulose [98] or a phospholipid nanogel [26,50]. After optimization of several on-line reaction parameters, such as the length of the enzyme plug, enzyme concentration, and incubation time, the on-line sequencing of the terminal residues of complex N-glycans was achieved [26,50,98]. …”

Section: Emerging Techniques and Future Directionsmentioning

confidence: 99%

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Advances in enzyme substrate analysis with capillary electrophoresis

Gattu

Crihfield

et al. 2018

Methods

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Capillary electrophoresis provides a rapid, cost-effective platform for enzyme and substrate characterization. The high resolution achievable by capillary electrophoresis enables the analysis of substrates and products that are indistinguishable by spectroscopic techniques alone, while the small volume requirement enables analysis of enzymes or substrates in limited supply. Furthermore, the compatibility of capillary electrophoresis with various detectors makes it suitable for KM determinations ranging from nanomolar to millimolar concentrations. Capillary electrophoresis fundamentals are discussed with an emphasis on the separation mechanisms relevant to evaluate sets of substrate and product that are charged, neutral, and even chiral. The basic principles of Michaelis-Menten determinations are reviewed and the process of translating capillary electrophoresis electropherograms into a Michaelis-Menten curve is outlined. The conditions that must be optimized in order to couple off-line and on-line enzyme reactions with capillary electrophoresis separations, such as incubation time, buffer pH and ionic strength, and temperature, are examined to provide insight into how the techniques can be best utilized. The application of capillary electrophoresis to quantify enzyme inhibition, in the form of KI or IC50 is detailed. The concept and implementation of the immobilized enzyme reactor is described as a means to increase enzyme stability and reusability, as well as a powerful tool for screening enzyme substrates and inhibitors. Emerging techniques focused on applying capillary electrophoresis as a rapid assay to obtain structural identification or sequence information about a substrate and in-line digestions of peptides and proteins coupled to mass spectrometry analyses are highlighted.

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“…For the analysis of glycoprotein glycans, an on‐line plug‐plug kinetic method has been developed . In this study, exoglycosidase digestion by several enzymes was used as a profiling tool of the glycans.…”

Section: On‐line Enzyme Reaction Studiesmentioning

confidence: 99%

Advances in Capillary Electrophoretically Mediated Microanalysis for On‐line Enzymatic and Derivatization Reactions

et al. 2017

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This review summarizes recent developments, applications, and innovations of capillary electrophoretically mediated microanalysis methods. As a follow up of an earlier review, it covers the literature from early 2015 to early 2017. This article is divided into three parts. In the first part, different types of mixing procedures and applications of enzyme mediated microanalysis are discussed; the second part summarizes immobilized enzyme reactors (IMERs), while the third part deals with recent advances in on-line derivatization reactions.

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Plug-plug kinetic capillary electrophoresis for in-capillary exoglycosidase digestion as a profiling tool for the analysis of glycoprotein glycans

Cited by 20 publications

References 35 publications

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Recent developments in capillary and microchip electroseparations of peptides (2015–mid 2017)

Advances in enzyme substrate analysis with capillary electrophoresis

Advances in Capillary Electrophoretically Mediated Microanalysis for On‐line Enzymatic and Derivatization Reactions

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