Glycoform analysis of alpha1-acid glycoprotein based on capillary electrophoresis and electrophoretic injection

Chenhua Zhang, Cong Bi, William Clarke, David S. Hage

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

A method based on capillary electrophoresis (CE) with electrophoretic injection and absorbance detection was developed for the direct analysis of AGP glycoforms in human serum. Electrophoretic injection of AGP was performed in the reversed-polarity mode of CE with a capillary coated with poly(ethylene oxide) and that had minimal electroosmotic flow. This situation created an essentially stationary interface between the sample and running buffer during injection and sample stacking. This approach allowed an 11,000-fold increase in sample loading for a 5 min injection versus hydrodynamic injection and without introducing any significant levels of extra band-broadening. This method was used with sample pretreatment methods based on acid precipitation and desalting to examine AGP glycoforms in only 65 μL of serum. A limit of detection of 2.1–11.3 nM was obtained for the major AGP glycoform bands in serum, and the sample pretreatment method gave a recovery of 72.3–80.9% for these glycoforms. The precision for the migration times was ± 0.08–0.13% and the precision for the peak areas was ± 0.34–1.18% when using serum samples and an internal standard. This method was used for both normal pooled serum and serum from individuals with systemic lupus erythematosus. Results were obtained in a separation time of 25 min and allowed the comparison of up to eleven glycoform bands in these samples. A similar approach may be useful in examining additional glycoproteins in serum or other types of biological samples.

Original languageEnglish (US)
Pages (from-to)114-122
Number of pages9
JournalJournal of Chromatography A
Volume1523
DOIs
StatePublished - Nov 10 2017

Fingerprint

Capillary electrophoresis
Capillary Electrophoresis
Glycoproteins
Salt removal
Injections
Acids
Polyethylene oxides
Serum
Buffers
Hydrodynamics
Recovery
Electroosmosis
Ethylene Oxide
Systemic Lupus Erythematosus
Limit of Detection

Keywords

  • Alpha-acid glycoprotein
  • Capillary electrophoresis
  • Electrophoretic injection
  • Glycoform analysis
  • Sample stacking
  • Systemic lupus erythematosus

ASJC Scopus subject areas

  • Analytical Chemistry
  • Organic Chemistry
  • Biochemistry
  • Medicine(all)

Cite this

Glycoform analysis of alpha1-acid glycoprotein based on capillary electrophoresis and electrophoretic injection. / Zhang, Chenhua; Bi, Cong; Clarke, William; Hage, David S.

In: Journal of Chromatography A, Vol. 1523, 10.11.2017, p. 114-122.

Research output: Contribution to journalArticle

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