Abnormal Hemoglobins: Laboratory Methods

Wajcman, Henri; Préhu, Claude; Bardakdjian‐Michau, Josiane; Promé, Danièlle; Riou, Jean; Godart, Christian; Mathis, Mireille; Hurtrel, Didier; Galactéros, Frédéric

doi:10.1081/hem-100104026

Cited by 78 publications

(54 citation statements)

References 21 publications

(15 reference statements)

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“…The separation of the different globin chain fractions was done by reversed phase liquid chromatography (RP-LC) and spectral analysis, as previously described. 18 RP-LC analyses were performed on a C4 Uptisphere (silica beads 5 mm; average pore size 300 Å) (Interchim, Montluçon, France) (4.6 × 250 mm). Isolated globin-chain fractions were identified by electrospray ionization-mass spectrometry, as previously described.…”

Section: Hemoglobin Composition Of Mature Cultured Red Blood Cells Amentioning

confidence: 99%

Red blood cell generation from human induced pluripotent stem cells: perspectives for transfusion medicine

Lapillonne¹,

Kobari²,

Mazurier³

et al. 2010

Haematologica

215

253

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The online version of this article has a a Supplementary Appendix. BackgroundEx vivo manufacture of red blood cells from stem cells is a potential means to ensure an adequate and safe supply of blood cell products. Advances in somatic cell reprogramming of human induced pluripotent stem cells have opened the door to generating specific cells for cell therapy. Human induced pluripotent stem cells represent a potentially unlimited source of stem cells for erythroid generation for transfusion medicine. Design and MethodsWe characterized the erythroid differentiation and maturation of human induced pluripotent stem cell lines obtained from human fetal (IMR90) and adult fibroblasts (FD-136) compared to those of a human embryonic stem cell line (H1). Our protocol comprises two steps: (i) differentiation of human induced pluripotent stem cells by formation of embryoid bodies with indispensable conditioning in the presence of cytokines and human plasma to obtain early erythroid commitment, and (ii) differentiation/maturation to the stage of cultured red blood cells in the presence of cytokines. The protocol dispenses with major constraints such as an obligatory passage through a hematopoietic progenitor, co-culture on a cellular stroma and use of proteins of animal origin. ResultsWe report for the first time the complete differentiation of human induced pluripotent stem cells into definitive erythrocytes capable of maturation up to enucleated red blood cells containing fetal hemoglobin in a functional tetrameric form. ConclusionsRed blood cells generated from human induced pluripotent stem cells pave the way for future development of allogeneic transfusion products. This could be done by banking a very limited number of red cell phenotype combinations enabling the safe transfusion of a great number of immunized patients. © F e r r a t a S t o r t i F o u n d a t i o n

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Section: Hemoglobin Composition Of Mature Cultured Red Blood Cells Amentioning

confidence: 99%

Red blood cell generation from human induced pluripotent stem cells: perspectives for transfusion medicine

Lapillonne¹,

Kobari²,

Mazurier³

et al. 2010

Haematologica

215

253

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“…Routine Hb analysis was performed by isoelectric focusing (IEF), Triton X-100Àurea polyacrylamide gel electrophoresis (PAGE), and highperformance liquid chromatography (HPLC) as described elsewhere [10]. The isopropyl alcohol stability test was carried out as described elsewhere [10]. Oxygen equilibrium curves were determined using a Hemox-Analyzer (TCS, New Hope, PA) [12].…”

Section: Methodsmentioning

confidence: 99%

New α2 globin chain variant with low oxygen affinity affecting the N‐terminal residue and leading to N‐acetylation [Hb Lyon‐Bron α 1(NA1)Val→Ac‐Ala]

Lacan¹,

Souillet²,

Aubry³

et al. 2002

American J Hematol

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Hemoglobin Lyon-Bron was found in two members of a family of German ascent presenting with a moderate normocytic anemia. In this alpha2 globin variant, the N-terminal valine of the chain was replaced by an alanine. Electrospray mass spectrometry of the a chain showed that, as normally, the initiator methionine was cleaved during globin processing but that the Na-terminal group was totally acetylated. This resulted in structural modi®cations of a region crucial for oxygen binding. As a consequence, hemoglobin Lyon-Bron displayed both a reduced chloride effect and a decreased oxygen af®nity, this last point explaining the apparent anemia. Am.

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“…Isoelectric focusing or high-performance liquid chromatography (HPLC), the most commonly used tests, identify most structurally abnormal Hbs (Wajcman et al 2001). In this way, many benign Hb variants are discovered incidentally.…”

Section: Laboratory Testing For Hb Variantsmentioning

confidence: 99%

“…As stated, "the primary reason for establishing the diagnosis [of M Hbs] is to prevent iatrogenic misadventures that might arise under the mistaken impression that the patient has a cardiac or pulmonary disorder" (Bunn and Forget 1986). Most Hb variants with altered O 2 affinity are relatively simple to diagnose through history, physical examination, and laboratory testing (Wajcman et al 2001;Wajcman and Moradkhani 2011).…”

Section: Identification Of Hb Variants and Their Clinical Implicationsmentioning

confidence: 99%

“…Specialized testing. Mass spectrometry analysis of patient hemolysate and DNA sequencing of globin genes are specialized confirmatory tests to identify amino acid and nucleotide alterations associated with suspected Hb variants (Wajcman et al 2001;Wajcman and Moradkhani 2011). DNA sequencing may readily elucidate the presence of an Hb variant.…”

Section: Laboratory Testing For Hb Variantsmentioning

confidence: 99%

See 1 more Smart Citation

Hemoglobin Variants: Biochemical Properties and Clinical Correlates

Thom

Dickson

Gell

et al. 2013

Cold Spring Harbor Perspectives in Medicine

225

182

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Diseases affecting hemoglobin synthesis and function are extremely common worldwide. More than 1000 naturally occurring human hemoglobin variants with single amino acid substitutions throughout the molecule have been discovered, mainly through their clinical and/or laboratory manifestations. These variants alter hemoglobin structure and biochemical properties with physiological effects ranging from insignificant to severe. Studies of these mutations in patients and in the laboratory have produced a wealth of information on hemoglobin biochemistry and biology with significant implications for hematology practice. More generally, landmark studies of hemoglobin performed over the past 60 years have established important paradigms for the disciplines of structural biology, genetics, biochemistry, and medicine. Here we review the major classes of hemoglobin variants, emphasizing general concepts and illustrative examples.

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Abnormal Hemoglobins: Laboratory Methods

Cited by 78 publications

References 21 publications

Red blood cell generation from human induced pluripotent stem cells: perspectives for transfusion medicine

Red blood cell generation from human induced pluripotent stem cells: perspectives for transfusion medicine

New α2 globin chain variant with low oxygen affinity affecting the N‐terminal residue and leading to N‐acetylation [Hb Lyon‐Bron α 1(NA1)Val→Ac‐Ala]

Hemoglobin Variants: Biochemical Properties and Clinical Correlates

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