High‐resolution capillary zone electrophoresis and mass spectrometry for distinction of undersialylated and hypoglycosylated transferrin glycoforms in body fluids

Abstract

High‐resolution capillary zone electrophoresis is used to distinguish transferrin glycoforms present in human serum, cerebrospinal fluid, and serum treated with neuraminidase and N‐glycosidase F. The obtained data are compared to mass spectrometry data from the literature. The main focus is on the analysis of the various asialo‐transferrin, monosialo‐transferrin, and disialo‐transferrin molecules found in these samples. The features of capillary zone electrophoresis and mass spectrometry are reviewed and highlighted in the context of the analysis of undersialylated and hypoglycosylated transferrin molecules. High‐resolution capillary zone electrophoresis represents an effective tool to assess the diversity of transferrin patterns whereas mass spectrometry is the method of choice to elucidate structural identification about the glycoforms. Hypoglycosylated transferrin glycoforms present in sera of alcohol abusers and normal subjects are structurally identical to those in sera of patients with a congenital disorder of glycosylation type I. Asialo‐transferrin, monosialo‐transferrin and disialo‐transferrin observed in sera of patients with a type II congenital disorder of glycosylation or a hemolytic uremic syndrome, in cerebrospinal fluid and after treatment of serum with neuraminidase are undersialylated transferrin glycoforms with two N‐glycans of varying structure. Undersialylated disialo‐transferrin is also observed in sera with high levels of trisialo‐transferrin.

Effect of nanocellulose polymorphism on electrochemical analytical performance in hybrid nanocomposites with non-oxidized single-walled carbon nanotubes

“…Another glycoprotein of high significance used as disease biomarker, transferrin [ 35 ], was analyzed using the same methodology (see Fig. S4 ).…”

Capillary electrophoresis of glycoproteins

“…Hypoglycosylated Tf forms were observed in serum samples from CDG type I patients while undersialylated Tf glycoforms were seen in CDG type II samples [54]. Analysis of these two types of Tf glycoforms by CE-UV and CE-MS have recently been reviewed [219]. Detection of Tf isoforms was possible in other body fluids besides serum, namely 2-Tf, an asialo-TF, was determined in cerebrospinal fluid (CSF), which is indicative of CSF leakage [85].…”

Structural Elucidation of β1- and β2-Transferrin Using Microprobe-Capture In-Emitter Elution and High-Resolution Mass Spectrometry

“…It was found that the N-glycans on the major serum-type Tf consisted of bi-antennary oligosaccharide chains with sialylated terminals, and those on the brain-type Tf are desialylated, or more specifically, unsialylated (asialotransferrin) (15,16,18,20). It was hypothesized that the major brain-type Tf was β 2 -Tf (21), but this hypothesis has not been proved.…”