Electrophoretic characteristics of monoclonal immunoglobulin G of different subclasses

Vu, Dinh-Hao; Schneider, Philippe; Tissot, Jean‐Daniel

doi:10.1016/s0378-4347(01)00384-x

Cited by 11 publications

(11 citation statements)

References 43 publications

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“…Analysis of monoclonal gammopathies [45,46] and characterization of cryoproteins [47][48][49] using 2-DE has been shown to be useful in selected difficult clinical situations. Using 2-DE analyses and MS, we also recently identified a new peptide, called hSpa or CD5L [50].…”

Section: Clinical Applications -Biomarker Identificationmentioning

confidence: 99%

Recent advances in blood‐related proteomics

et al. 2005

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Service régional vaudois de transfusion sanguine, Lausanne, SwitzerlandBlood is divided in two compartments, namely, plasma and cells. The latter contain red blood cells, leukocytes, and platelets. From a descriptive medical discipline, hematology has evolved towards a pioneering discipline where molecular biology has permitted the development of prognostic and diagnostic indicators for disease. The recent advance in MS and protein separation now allows similar progress in the analysis of proteins. Proteomics offers great promise for the study of proteins in plasma/serum, indeed a number of proteomics databases for plasma/ serum have been established. This is a very complex body fluid containing lipids, carbohydrates, amino acids, vitamins, nucleic acids, hormones, and proteins. About 1500 different proteins have recently been identified, and a number of potential new markers of diseases have been characterized. Here, examples of the enormous promise of plasma/serum proteomic analysis for diagnostic/prognostic markers and information on disease mechanism are given. Within the blood are also a large number of different blood cell types that potentially hold similar information. Proteomics of red blood cells, until now, has not improved our knowledge of these cells, in contrast to the major progresses achieved while studying platelets and leukocytes. In the future, proteomics will change several aspects of hematology.

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Section: Clinical Applications -Biomarker Identificationmentioning

confidence: 99%

Recent advances in blood‐related proteomics

et al. 2005

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“…To validate the utility of the high-sensitive 3-DE method, we compared it to the method described by Vu et al [33]. The recovery of M-proteins with the high-sensitive method was clearly higher than with the method of Vu et al (Figure 2A).…”

Section: Discussionmentioning

confidence: 96%

“…In order to validate the utility of the high-sensitive 3-DE method, we first compared the percentage of M-protein recovered in the agarose gel electrophoresis with the high-sensitive method and the original method developed by Vu et al [33]. As shown in Figure 2A, recovery of M-protein was significantly higher with the high-sensitive method than with the original method.…”

Section: Validation Of the Utility Of The High-sensitive 3-de Methodsmentioning

confidence: 99%

“…Samples were also analyzed using the original 7 3-DE described by Vu et al [33] to validate the utility of the high-sensitive 3-DE.…”

Section: High-sensitive 3-dementioning

confidence: 99%

“…M-proteins were separated using agarose gel electrophoresis, extracted from the gel film using solutions containing SDS, and subsequently subjected to 2-DE [30; 31; 32]. Vu et al developed methods for extracting M-proteins from commercially available agarose film using centrifugation rather than solutions, and they termed this technique three-dimensional gel electrophoresis (3-DE) [33]. However, this method has not been optimized, especially with regard to the M-protein extraction from the first dimension.…”

Section: Introductionmentioning

confidence: 99%

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Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity

Nakano

Tamura

Otuka

et al. 2013

Analytical Biochemistry

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Three-dimensional gel electrophoresis (3-DE), which combines agarose gel electrophoresis and IEF/SDS-PAGE, was developed to characterize monoclonal proteins (M-proteins). However, original 3-DE method has not been optimized and its specificity has not been demonstrated. The main goal of this study was to optimize the procedure of 3-DE and then compare it with 2-DE. We developed a high-sensitive 3DE method in which M-proteins extracted from first dimension, agarose gel, by diffusing into 150mM NaCl and the recovery of M-proteins was 90.6%. To validate the utility of the high-sensitive 3DE, we compared it with the original 3-DE method. We found that high-sensitive 3DE provided for greater M-proteins recovery and was more effective in terms of detecting spots on SDS-PAGE gels than the original 3-DE. Moreover, high-sensitive 3-DE separates residual normal IgG from M-proteins which could not be done by 2-DE. Applying the high-sensitive 3-DE to clinical samples, we found that the characteristics of M-proteins vary tremendously between individuals. We believe that our high-sensitive 3-DE method described here will prove useful in further studies of the heterogeneity of M-proteins.

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The immunoglobulinopathies: From physiopathology to diagnosis

Tissot

Aubert

et al. 2002

Proteomics

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The diversity of immunoglobulins (Igs) results mainly from recombinations of numerous genes within the heavy (V(Heavy), D, and J(Heavy)) and within the light (V(Light), J(Light)) chain gene loci, and from somatic hypermutations occurring during the immune response of B-cells. Igs production is controlled by complex cellular and humoral mechanisms. Plasma of healthy individuals contains polyclonal Igs. Clonal expansion of cells producing antigen-specific Igs may result from physiological as well as from pathological immune events. Exquisitely sensitive and specific molecular biology techniques have been used to evaluate the clonal diversity of cells producing antigen-specific Ig. However, the application of such techniques is hampered by the necessity to collect the totality of antigen-specific B-cells for subsequent analysis, which is impossible to perform routinely in humans. In addition, these techniques do not provide quantitative information about the concentration of the circulating Igs. It is therefore necessary to use tools allowing study of the quantity of the circulating Igs, and more particularly to detect overproduction of a single homogeneous Ig resulting from the expansion of a B-cell clone secreting Igs. Here, we review the mechanisms of B-cell differentiation and Ig synthesis, discuss the diseases associated with clonal Ig production and review the methods available in the clinical laboratory for Ig analysis.

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Electrophoretic characteristics of monoclonal immunoglobulin G of different subclasses

Cited by 11 publications

References 43 publications

Recent advances in blood‐related proteomics

Recent advances in blood‐related proteomics

Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity

The immunoglobulinopathies: From physiopathology to diagnosis

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