Hemoglobin Variant Analysis of Whole Blood and Dried Blood Spots by MS

Abstract

MS allows for the unequivocal diagnosis of hemoglobin variants, or hemoglobinopathies. Hemoglobinopathies are the most common inherited disorder and there is a need for rapid detection of clinically significant variants, such as sickle hemoglobin, which is responsible for sickle cell disease. In this review, we describe the development of MS approaches for the determination of hemoglobin variants from both whole blood samples and dried blood spots. MS approaches that are suitable for population screening are discussed, as are recent advances in direct surface analysis of dried blood spots.

Capillary Zone Electrophoresis-Top-Down Tandem Mass Spectrometry for In-Depth Characterization of Hemoglobin Proteoforms in Clinical and Veterinary Samples

“…The use of dried blood spots (DBS) is an emerging technique in the clinical context with several advantages such as the low sample amounts needed, low costs and easy handling, storage, and shipping . DBS is routinely used in newborn screening for genetic diseases such as phenylketonuria and sickle cell disease . The application of DBS samples for Hb analysis in general has been reviewed, summarizing the applications and advantages of the technique …”

“…DBS is routinely used in newborn screening for genetic diseases such as phenylketonuria and sickle cell disease . The application of DBS samples for Hb analysis in general has been reviewed, summarizing the applications and advantages of the technique …”

“…32 DBS is routinely used in newborn screening for genetic diseases such as phenylketonuria 33 and sickle cell disease. 34 The application of DBS samples for Hb analysis in general has been reviewed, summarizing the applications and advantages of the technique. 34 The aim of this study was to develop a versatile screening method for the joint characterization of various Hb proteoforms (including both, sequence variants as well as posttranslationally modified species) from clinical human and veterinary DBS samples.…”

Smart proteolysis samplers for pre‐lab bottom‐up protein analysis – Performance of on‐paper digestion compared to conventional digestion

“…Mass spectrometry offers the possibility to perform targeted protein analysis as well as shotgun proteomic analysis. For targeted analysis one of the first reports is the MS-based determination of hemoglobin F-Mauritius from DBS [13] which was followed by a long list of other hemoglobin variants determined using MS with bottom-up and top-down approaches [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31]. Not only hemoglobin was determined in this way, but also targeted determination of other proteins like human chorionic gonadotropin [32,33], insulin [34,35] and apolipoprotein A-I, apolipoprotein B [36] as well as multiplex strategies using multiple reaction monitoring (MRM) were reported [37][38][39][40].…”

Identification of hemoglobin variants by top-down mass spectrometry using selected diagnostic product ions

“…Several approaches using top-down MS were also developed for the identification and characterization of common Hb variants [5,[11][12][13]. Compared to bottom-up methods, intact protein approaches have, in principle, the advantage to reduce the number of sample preparation steps and decrease or eliminate sample preparation time [14]. Cooper and co-workers showed recently the possibility to detect Hb variant with a mass shift less than 1 Da by direct surface sampling top-down high-resolution MS [15].…”