Discrimination of glycoproteins via two‐color laser‐induced fluorescence detection coupled with postcolumn derivatization in capillary electrophoresis

Tabara, Ayumi; Kaneta, Takashi

doi:10.1002/elps.201300149

Cited by 4 publications

(6 citation statements)

References 41 publications

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“…The post‐column reactors developed by our group are shown in Figure . One consists of a laser‐drilled capillary which has a small hole with a diameter similar to the inner diameter of the capillary, and another uses the sheath flow of a labeling reagent . Initially, the post‐column reactor consisting of a laser‐drilled capillary was employed in fluorescence detection using an ultraviolet light emitting diode (UV‐LED) to detect amino acids derivatized by o ‐phthalaldehyde (OPA), which is a fluorogenic reagent used to form fluorescent derivatives via reaction with primary amines .…”

Section: Overview Of Ce‐lifmentioning

confidence: 99%

“…A post‐column reactor using sheath flow was applied to two‐color LIF to discriminate a glycosylated protein from an unglycosylated protein ,. We employed two lasers emitting at 450 (diode laser) and 532 nm (diode laser‐pumped Nd : YAG laser) that were focused on a capillary in the same position in order to simultaneously record two electropherograms.…”

Section: Overview Of Ce‐lifmentioning

confidence: 99%

“…Via use of the proposed principle, we differentiated a glycosylated protein (thyroglobulin) from an unglycosylated protein (albumin) in the presence of Rh‐ConA. The separation of thyroglobulin and albumin is shown in Figure . In Figure (a), the sample contained two proteins that could not be resolved in the BGE consisting of 50 mM of borate buffer (pH 9.2), which resulted in a single peak.…”

Section: Overview Of Ce‐lifmentioning

confidence: 99%

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Laser‐Induced Fluorometry for Capillary Electrophoresis

Kaneta

2018

The Chemical Record

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Laser-induced fluorometry (LIF) has achieved the detection of single molecules, which ranks it among the most sensitive of detection techniques, whereas capillary electrophoresis (CE) is known as a powerful separation method with resolution that is beyond the reach of many other types of chromatography. Therefore, a coupling of LIF with CE has established an unrivaled analytical technique in terms of sensitivity and resolution. CE-LIF has demonstrated excellent performance in bioanalytical chemistry for the high-resolution separation and highly sensitive detection of DNAs, proteins, and small bioactive molecules. This review describes the CE-LIF methods developed by the author's group that include indirect and direct detection using diode lasers, post-column derivatization, and Hadamard transformation, as well as applications to the binding assays of specific DNA immunoassays of proteins and to the determination of anticancer drugs.

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Section: Overview Of Ce‐lifmentioning

confidence: 99%

Section: Overview Of Ce‐lifmentioning

confidence: 99%

Section: Overview Of Ce‐lifmentioning

confidence: 99%

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Laser‐Induced Fluorometry for Capillary Electrophoresis

Kaneta

2018

The Chemical Record

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“…Three articles were from this mode [53][54][55]. Abdul Keyon et al analysed paralytic shellfish toxins by post-capillary oxidation using droplet-microfluidics [53].…”

Section: Post-capillary Derivatisationmentioning

confidence: 99%

“…Pre‐capillary oxidation of paralytic shellfish toxins can result in multiple and identical derivatives from different toxins which complicates the analysis. Tabara and Kaneta used a post‐capillary derivatisation chamber to analyse glycoproteins from HSA . A T‐piece connected a 50 μm I.D.…”

Section: Derivatisation Modesmentioning

confidence: 99%

Derivatisation for separation and detection in capillary electrophoresis (2012–2015)

Wuethrich

Quirino

2015

Electrophoresis

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Derivatisation is a well-established and mature form of sample preparation for CE. The modification of the analyte can cause superior analysis characteristics such as better sensitivity and selectivity, however, derivatisation of the analyte introduces an additional step into the analytical workflow. This review covers articles from January 2012 to January 2015 on derivatisation in CE. The main sections are on the derivatisation modes (i.e. pre-capillary, in-line, in-capillary and post-capillary), separation and detection modes (i.e. LIF and others). LIF is discussed in more detail since this detection mode was most prevalent. A table of the common labelling agents and wavelengths for excitation and emission and the common derivatisation reactions are included. In addition, a comprehensive table which summarises all research articles is provided. This review is suitable for analytical chemists as a guide for 'how to get started' with derivatisation for separation and detection in CE.

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Discrimination of Glycoproteins from Unglycosylated Proteins in Capillary Electrophoresis: Two-Color LIF Detection Coupled with Post-column Derivatization

Kaneta

2016

Methods in Molecular Biology

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Glycosylation is one of the most important posttranslational modifications (PTMs) which lead to the functionalization of proteins. Here, we describe one method for discriminating glycosylated proteins from unglycosylated ones in their mixture sample by capillary electrophoretic separation and two-color laser-induced fluorescence detection coupled with post-column derivatization. Two lasers emitting at 450 and 532 nm permit the detection of amino groups of proteins derivatized by naphthalene-2,3-dicarboxaldehyde and a fluorescently labeled lectin, tetramethylrhodamine-labeled concanavalin A (Rh-Con A), respectively. When a protein mixture react with Rh-Con A, the glycoproteins bound with Rh-Con A exhibit signals at the same migration time in two electropherograms obtained by 450- and 532-nm lasers whereas unbound proteins show a signal only in the electropherogram of the 450-nm laser. So, when one protein is glycosylated it is detected at the same migration time in the electropherograms obtained by two lasers.

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Discrimination of glycoproteins via two‐color laser‐induced fluorescence detection coupled with postcolumn derivatization in capillary electrophoresis

Abstract: 2Here, we report a novel method consisting of capillary electrophoretic separation followed by two-color laser-induced fluorescence detection with postcolumn derivatization.

Cited by 4 publications

References 41 publications

Laser‐Induced Fluorometry for Capillary Electrophoresis

Laser‐Induced Fluorometry for Capillary Electrophoresis

Derivatisation for separation and detection in capillary electrophoresis (2012–2015)

Discrimination of Glycoproteins from Unglycosylated Proteins in Capillary Electrophoresis: Two-Color LIF Detection Coupled with Post-column Derivatization

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