Stolz, Alexander and Hedeland, Ylva and Salzer, Liesa and Roemer, Jennifer and Heiene, Reidun and Leclercq, Laurent and Cottet, Herve and Bergquist, Jonas and Neusuess, Christian (2020) Capillary Zone Electrophoresis-Top-Down Tandem Mass Spectrometry for In-Depth Characterization of Hemoglobin Proteoforms in Clinical and Veterinary Samples. ANALYTICAL CHEMISTRY, 92 (15). pp. 10531-10539. ISSN 0003-2700, 1520-6882
Full text not available from this repository. (Request a copy)Abstract
Hemoglobin (Hb) constitutes an important protein in clinical diagnostics-both in humans and animals. Among the high number of sequence variants, some can cause severe diseases. Moreover, chemical modifications such as glycation and carbamylation serve as important biomarkers for conditions such as diabetes and kidney diseases. In clinical routine analysis of glycated Hb, sequence variants or other Hb proteoforms can cause interference, resulting in wrong quantification results. We present a versatile and flexible capillary zone electrophoresis-mass spectrometry screening method for Hb proteoforms including sequence variants and modified species extracted from dried blood spot (DBS) samples with virtually no sample preparation. High separation power was achieved by application of a 5-layers successive multiple ionic polymer layers-coated capillary, enabling separation of positional isomers of glycated alpha- and beta-chains on the intact level. Quantification of glycated Hb was in good correlation with the results obtained in a clinical routine method. Identification and characterization of known and unknown proteoforms was performed by fragmentation of intact precursor ions. N-Terminal and lysine glycation could be identified on the alpha- and beta-chain, respectively. The versatility of the method was demonstrated by application to dog and cat DBS samples. We discovered a putative new sequence variant of the beta-chain in dog (T38 -> A). The presented method enables separation, characterization, and quantification of intact proteoforms, including positional isomers of glycated species in a single run. Combined with the simple sample preparation, our method represents a valuable tool to be used for deeper characterization of clinical and veterinary samples.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | DRIED BLOOD SPOTS; CARBAMYLATED HEMOGLOBIN; GLYCATION; HPLC; IDENTIFICATION; HEMODIALYSIS; PROTEOMICS; VARIANTS; PROTEINS; HBA(1C); |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 17 Mar 2021 09:01 |
| Last Modified: | 17 Mar 2021 09:01 |
| URI: | https://pred.uni-regensburg.de/id/eprint/44056 |
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