Ultra-Fast Analysis of Hemoglobin Isoforms Using Capillary Electrophoresis

仪器资料 · 发表于 2019-6-5 17:09:45

Ultra-Fast Analysis of Hemoglobin Isoforms Using Capillary Electrophoresis Coupled to Mass Spectrometry and Ultraviolet Photodissociation

ABSTRACT WORKFLOW
Capillary zone electrophoresis (CZE) coupled to Mass Spectrometry (MS) is a powerful
technique to assess the structure and identity of individual bioanalytes based on their size-tocharge
ratio, including intact proteins over a wide molecular weight range. Therefore, CZE-MS
is a promising approach for top down proteomics, because it is suited to separate proteins with
post-translational modifications (PTMs) or degradations that affect the charge state of targeted
molecules, including deamidation, glycosylation, and phosphorylation. CZE-MS is a promising
approach for hemoglobin profiling in the clinical research field to routinely detect abnormal
proteoforms of hemoglobin. Here, we are evaluating a CZE-MS system using a newly
developed and commercially available sheath flow interface1 that sustains a stable
electrospray into a Thermo Scientific Orbitrap Fusion™ Lumos™ Tribrid™ mass spectrometer,
as well as comparing various fragmentation methods to obtain best amino acid sequence
coverage via top down proteomic analysis.