Characterization of human transferrin glycoforms by capillary electrophoresis and electrospray ionization mass spectrometry

Sanz‐Nebot, Victoria; Gonzalez, Pascual; Toro, I.; Ribes, Antònia; Barbosa, J.

doi:10.1016/s1570-0232(02)00442-7

Cited by 29 publications

(32 citation statements)

References 28 publications

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“…Neuraminidase cleaves the terminal sialic acid residues and thus, the peak of the tetrasialylated isoform decreases while new peaks containing 0, 1, 2, and 3 sialic acids appear at lower migration times. This methodology was followed in a previous article [25] for Tf glycoform assignment. Here, the mobility scale conversion and the further CE-ESI-MS coupling allow the unequivocal identification of the observed glycoforms.…”

Section: Cdt Detection In Serum Samples By Ce-uvmentioning

confidence: 99%

“…Figure 3 shows several electrophoretic profiles obtained by CE-UV and converted to the m e scale for different serum samples. Figures 3a and b show the glycoform profile for two control sera from healthy individuals whose main peak corresponds to the tetrasialoform obtained at m e 29.5610 25 . Tri-, penta-, and hexasialoforms have been observed in less intensity.…”

Section: Cdt Detection In Serum Samples By Ce-uvmentioning

confidence: 99%

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Characterization of transferrin glycoforms in human serum by CE‐UV and CE‐ESI‐MS

et al. 2007

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Human transferrin (Tf) is a model glycoprotein for congenital disorders of glycosylation (CDG) diagnosis. In the last few years, new CE-UV methods for intact Tf glycoforms analysis have been developed using nonvolatile BGEs and organic modifiers. However, the use of these BGEs does not allow the coupling of these procedures with electrospray MS (ESI-MS). In this study, a new CE-UV separation method of Tf glycoforms is developed, using a double-layer stable coating and a volatile BGE based on ammonium acetate. The separation method is optimized using standard Tf and their potential is demonstrated applying the method to the analysis of sera Tf from healthy individuals and CDG patients. The CE-UV separation method has been coupled to ESI-MS detection. Main parameters such as sheath liquid composition are optimized in order to obtain a good sensitivity. The CE-ESI-MS method has also been used in serum samples obtaining the separation of the different proteins present in serum and partial separation of Tf glycoforms. Different mass spectra and deconvoluted molecular masses were obtained for each sialoform, allowing unequivocal glycoform identification.

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Section: Cdt Detection In Serum Samples By Ce-uvmentioning

confidence: 99%

Section: Cdt Detection In Serum Samples By Ce-uvmentioning

confidence: 99%

Characterization of transferrin glycoforms in human serum by CE‐UV and CE‐ESI‐MS

et al. 2007

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“…A means to close the CE electrical circuit for analyte separation while simultaneously providing an electrical potential to the spray tip is necessary. The common interfaces can be divided into three categories: sheath-flow 16,[18][19][20]24,27,32,37,38,40,43,47,50,[53][54][55][56]60,66 and sheathless 15,17,25,26,[28][29][30]34,36,44,45,48,49,61 and liquid-junction interfaces. [92][93][94] In the sheath-flow design, a coaxial sheath liquid, usually a hydroorganic mixture, mixes with the capillary eluent at the outlet end of the capillary.…”

Section: ·1 Esi-msmentioning

confidence: 99%

“…1 Moreover, it provides another window for optimizing the detection parameters and tuning selectivity. Sanz-Nebot et al, 53 in comparing the effect of methanol, 2-propanol and acetonitrile as an organic modifier in the sheath liquid, showed that the highest signal intensities for some peptide hormones were obtained using methanol. A study by Brenner-Weis et al 54 indicated that increasing the concentration of methanol in the sheath liquid favored the detection of dissociated myoglobin complexes over the intact species, and that even slight changes in the pH of the sheath liquid affected the stability of the complexes.…”

Section: ·1 Esi-msmentioning

confidence: 99%

“…In this study, significantly higher amounts of GnRH were found in serum and urine samples from a patient suffering from benign prostatic hyperplasia compared to a normal subject. Using CE-MS, Sanz-Nebot et al 53 compared the glycoforms of serum transferrin (Tf) in healthy individuals and Carbohydrate Deficient Glycoprotein Syndrome (CDGS) patients, their results indicating a marked variation in terms of the type and concentration of glycoforms between the two groups. Wittke et al 56 also used CE-MS to obtain patterns of proteins and peptides present in the urine of healthy individuals ("normal urine polypeptide pattern").…”

Section: ·3 Clinical Diagnosticsmentioning

confidence: 99%

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Recent Developments in Capillary Electrophoresis–Mass Spectrometry of Proteins and Peptides

Monton

Terabe

2005

ANAL. SCI.

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Many researchers have invested considerable efforts toward improving capillary electrophoresis (CE)-mass spectrometry (MS) systems so they can be applied better to standard analyses. This review highlights the developments in CE-MS of proteins and peptides over the last five years. It includes the developments in interfaces, sample-enrichment techniques, microfabricated devices, and some applications, largely in capillary zone electrophoresis (CZE), capillary isoelectric focusing (CIEF) and capillary isotachophoresis formats.

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Capillary zone electrophoresis for determination of carbohydrate‐deficient transferrin in human serum

et al. 2004

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Carbohydrate-deficient transferrin (CDT) is the most specific marker for diagnosis of chronic excessive alcohol consumption and includes the serum transferrin (Tf) isoforms with two or less sialic acid residues (di-, mono-, and asialo-Tf). To monitor serum CDT, we developed a capillary zone electrophoresis (CZE) method based on the dynamic capillary coating with diethylenetriamine (DETA). The separation was performed in a bare fused-silica capillary (50 microm ID, 57 cm in length), applying a voltage of 25 kV and a temperature of 40 degrees C. Using a 100 mmol/L borate buffer, pH 8.4 with 3 mmol/L DETA, the Tf isoforms (asialo- to pentasialo-Tf) were resolved within 16 min. Enzymatic cleavage of sialic acid residues with neuraminidase and immunosubtraction were used to identify CDT isoforms. The relative amount of CDT expressed as area % of disialo-Tf isoform related to the area of tetrasialo-Tf in 50 healthy donors (24 males and 26 females; aged 25-50 years) was 3.15 +/- 0.76% (mean +/- SD). The comparison between CDT values obtained by this CZE procedure and the "Axis-Shield %CDT" kit gave r = 0.644, p < 0.001 (n = 290). This easy to use and inexpensive CZE procedure could be an ideal tool to investigate CDT proteins for clinical or forensic purposes.

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Characterization of human transferrin glycoforms by capillary electrophoresis and electrospray ionization mass spectrometry

Cited by 29 publications

References 28 publications

Characterization of transferrin glycoforms in human serum by CE‐UV and CE‐ESI‐MS

Characterization of transferrin glycoforms in human serum by CE‐UV and CE‐ESI‐MS

Recent Developments in Capillary Electrophoresis–Mass Spectrometry of Proteins and Peptides

Capillary zone electrophoresis for determination of carbohydrate‐deficient transferrin in human serum

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