High Performance Liquid Cbromatographic Separation of Globin Chains on a Large-Pore C<sub>4</sub>Column

Shelton, J. C.; Shelton, J. R.; Schroeder, W. A.

doi:10.1080/01483918408068849

Cited by 431 publications

(118 citation statements)

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“…Freshly drawn blood was adjusted to a packed cell volume of 50% and incubated for i h at 370C in the presence of [3H]1-leucine under conditions previously detailed (27). The globin chains were separated by high performance liquid chromatography (HPLC) on large-pore C4 columns (Zydac Inc.) (28) and the radioactivity incorporated into a-and fl-globin chains was determined as previously described (27).…”

Section: Methodsmentioning

confidence: 99%

Molecular basis for nondeletion alpha-thalassemia in American blacks. Alpha 2(116GAG----UAG).

Liebhaber¹,

Coleman²,

Adams³

et al. 1987

J. Clin. Invest.

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Section: Methodsmentioning

confidence: 99%

Molecular basis for nondeletion alpha-thalassemia in American blacks. Alpha 2(116GAG----UAG).

Liebhaber¹,

Coleman²,

Adams³

et al. 1987

J. Clin. Invest.

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“…The hemoglobin solution (100 g) was used to determine the globin chain content by high pressure liquid chromatography on a Vydac large pore C 4 column (4.6 ϫ 250 mm; Grace Vydac, Hesperia, CA). Globin chains were resolved with a helium-degassed modified trifluoroacetic acid/ acetonitrile gradient: phase A (0.18% trifluoroacetic acid (w/v) in 36% acetonitrile) and phase B (0.18% trifluoroacetic acid (w/v) in 46% acetonitrile) (20,21). Globin chains were eluted by increasing mobile phase B from 33 to 37% over 15 min and, subsequently, to 70% over 35 min at a flow rate of 1 ml/min, and the elution profiles were followed by UV detection at 220 nm.…”

Section: Globin Chain Analysismentioning

confidence: 99%

Differential Regulatory and Compensatory Responses in Hematopoiesis/Erythropoiesis in α- and β-Globin Hemizygous Mice

Beauchemin¹,

Blouin

Trudel

2004

Journal of Biological Chemistry

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Characterization of hematopoiesis/erythropoiesis in thalassemias from multipotent primitive cells to mature erythrocytes is of fundamental importance and clinical relevance. We investigated this process in ␣-and ␤-globin hemizygous mice, lacking the two adult tandemly organized genes from either the ␣-or ␤-globin locus. Although both mice backcrossed on a homogeneous background exhibited similar reduced red blood cell (RBC) survival, ␤-globin hemizygous mice had less severe reticulocyte loss and globin chain imbalance, suggesting an apparently milder thalassemia than for ␣-globin hemizygous mice. In contrast, however, ␤-globin hemizygous mice displayed a more marked perturbation of hematologic parameters. Quantification of erythroid precursor subpopulations in marrow and spleen of ␤-globin hemizygous mice showed more severely impaired maturation from the basophilic to orthochromatophilic erythroblasts and substantial loss of these late precursors probably as a consequence of a greater susceptibility to an excess of free ␣-chain than ␤-chain. Hence, only erythroid precursors exhibiting stochastically moderate chain imbalance would escape death and mature to reticulocyte/RBC stage, leading to survival and minimal loss of reticulocytes in the ␤-globin hemizygous mice. Furthermore, in response to the ineffective erythropoiesis in ␤-globin hemizygous mice, a dynamic compensatory hematopoiesis was observed at earlier differentiation stage as evidenced by a significant increase of erythroid progenitors (erythroid colony-forming units ϳ100-fold) as well as of multipotent primitive cells (day 12 spleen colony-forming units ϳ7-fold). This early compensatory mechanism was less pronounced in ␣-globin hemizygous mice. The expansion of multipotent primitive and potentially stem cell populations, taken together with ineffective erythropoiesis and increased reticulocyte/RBC destruction could confer major cumulative advantage for gene targeting/bone marrow transplantation. Therefore, this study not only corroborated the strong potential effectiveness of transplantation for thalassemic hematopoietic therapy but also demonstrated the existence of a differential regulatory response for ␣-and ␤-thalassemia.

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“…When detected by the former procedure, HbF was quantified by the alkali denaturation method of Betke [14]. ␥ Chain typing in HbF was performed by HPLC [15]. LPLC was used to quantify HbA 2 [16].…”

Section: Hematological Studiesmentioning

confidence: 99%

Spectrum of β thalassemia mutations and HbF levels in the heterozygous Moroccan population

et al. 2003

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High Performance Liquid Cbromatographic Separation of Globin Chains on a Large-Pore C₄Column

Cited by 431 publications

References 5 publications

Molecular basis for nondeletion alpha-thalassemia in American blacks. Alpha 2(116GAG----UAG).

Molecular basis for nondeletion alpha-thalassemia in American blacks. Alpha 2(116GAG----UAG).

Differential Regulatory and Compensatory Responses in Hematopoiesis/Erythropoiesis in α- and β-Globin Hemizygous Mice

Spectrum of β thalassemia mutations and HbF levels in the heterozygous Moroccan population

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High Performance Liquid Cbromatographic Separation of Globin Chains on a Large-Pore C4Column

Cited by 431 publications

References 5 publications

Molecular basis for nondeletion alpha-thalassemia in American blacks. Alpha 2(116GAG----UAG).

Molecular basis for nondeletion alpha-thalassemia in American blacks. Alpha 2(116GAG----UAG).

Differential Regulatory and Compensatory Responses in Hematopoiesis/Erythropoiesis in α- and β-Globin Hemizygous Mice

Spectrum of β thalassemia mutations and HbF levels in the heterozygous Moroccan population

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Product

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High Performance Liquid Cbromatographic Separation of Globin Chains on a Large-Pore C₄Column