Performance Comparison of Capillary and Agarose Gel Electrophoresis for the Identification and Characterization of Monoclonal Immunoglobulins

McCudden, Christopher R.; Mathews, Stephanie; Hainsworth, Shirley A.; Chapman, John F.; Hammett‐Stabler, Catherine A.; Willis, Monte S.; Grenache, David G.

doi:10.1309/6kt8n49brnvvvbt1

Cited by 54 publications

(24 citation statements)

References 11 publications

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“…Unlike previous studies [9][10][11][12], which reported slightly superior sensitivity for CE compared to semiautomated AGE in detecting monoclonal immunoglobulins, our data shows that, at equal specificity, the fully automated AGE method (Interlab) is more sensitive than CE (Sebia), even when monoclonal proteins are present in low concentrations. It therefore appears that greater accuracy in the early detection of asymptomatic myeloma, as well as precise monitoring of malignant evolution and/or the efficacy of immunosuppressive treatment are now within reach.…”

contrasting

confidence: 91%

Comparison of the first fully automated agarose gel electrophoresis system with the Capillarys electrophoresis method for the identification of monoclonal immunoglobulins

et al. 2014

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Serum protein electrophoresis (SPE), used to identify monoclonal gammopathies, is generally performed using the "gold standard" agarose gel as migration support (AGE). However, despite being semi-automated, this electrophoresis technique remains labour-intensive, meaning analytical performance and throughput are limited. Hence fully automated capillary electrophoresis (CE), which enables rapid separation, has been adapted for use in clinical laboratories within the last decade [1][2][3][4][5]. Any qualitative abnormalities suggestive of monoclonal immunoglobulin detected by either AGE or CE are further analysed by immunofixation electrophoresis (IFE), to confirm the presence of monoclonal bands and to identify the type of monoclonal component (MC) [6].In order to increase throughput, a fully automated innovative AGE technique-Agarose Gel Easy Interlab G26 electrophoresis (Interlab, Rome, Italy)-has recently been launched, to be used with purpose-designed IFE kits.The aim of this study was to compare the AGE system with the Capillarys 2 ® CE equipment (Sebia, Evry, France) currently in use in our laboratory, to detect serum MCs subsequently confirmed, characterized and quantified by IFE on the same automated platform. In addition to method performance characteristic, the ease of use of the new system was analysed to evaluate its feasibility for real-world clinical scenarios.We analysed 300 consecutive serum specimens sent to our Clinical Pathology Laboratory for routine serum protein electrophoresis tests. These specimens were taken from patients hospitalized in Surgery (department connected to our Laboratory) during their pre-operative screening tests. All subjects provided informed consent prior to their participation, and the study was approved by the hospital Ethics Committee.CE was performed on Capillarys 2 ® [Capillarys Protein (E)6 kit] (Sebia), used for routine electrophoresis analysis in our laboratory, according to the manufacturer's instructions.The same sera were analysed on the same day using the new Agarose Gel electrophoresis system, also according to the manufacturer's instructions, in which human SPE performed at pH 8.9 yields five distinct, well-resolved zones or bands, each containing one or more different proteins. The patterns are examined visually for abnormalities and variations in the separated bands or zones, and densitometry is used for relative quantification of protein zones.Both sets of electrophoresis patterns were examined by two independent investigators for the presence (or suspicion) of MCs. IFE kits were used for confirmation, and to highlight any false positives or negatives, in samples in which one or both investigators detected apparent or evident MCs, as well as to identify and quantify the type of MC present.These new IFE kits have been designed for use with the AGE instrument under investigation, and feature ready-touse reagents, sample dilution phases, anti-serum dispensers, and blotting incubation. Acid violet is used as a highly sensitive staining solution, and the new e...

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contrasting

confidence: 91%

Comparison of the first fully automated agarose gel electrophoresis system with the Capillarys electrophoresis method for the identification of monoclonal immunoglobulins

et al. 2014

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“…81% (95% CI 70–92%) by Yang et al . but the characterized specimens in these studies did not contain a HCD (9–12). Yang et al .…”

Section: Discussionmentioning

confidence: 82%

Gamma heavy chain disease in a patient with rheumatoid arthritis – a laboratory evaluation

et al. 2012

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Introduction: Heavy chain diseases (HCD) are neoplastic proliferations of B cells which secrete truncated immunoglobulin heavy chains without associated light chains. Being rare and probably underdiagnosed diseases the aim of this report is to show an additional case of gamma heavy chain disease in a 48 year old female patient with rheumatoid arthritis focusing on the laboratory presentation. Materials and methods: Laboratory work-up included agarose gel electrophoresis (AGE), capillary zone electrophoresis (CZE), immunofixation and nephelometrically determined immunoglobulin and immunoglobulin subclasses of the patient's serum. Urine samples were also subjected to immunofixation and to a SDS-PAGE with consecutive immunoblot. Results: Nephelometrically measured elevated IgG concentrations were noted in combination with a decreased gamma globulin region and an increased beta globulin region on AGE. A definite monoclonal spike was not identified on AGE but at least suspected on CZE; finally serum and urine immunofixation demonstrated a monoclonal gamma heavy chain devoid of any corresponding light chains confirming the diagnosis of HCD. Analysis of the gamma heavy chain (HC) with means of SDS-PAGE revealed proteins of 40 kD and 80 kD most likely presenting a truncated HC in its monomeric and dimeric form and possibly leading to the failure of IgG-subclass typing with the applied IgG subclass antisera. Conclusion: This case report illustrates a new case of gamma HCD demonstrating variable laboratory manifestations and therefore the need for heightened awareness concerning this disease when confronted with abnormal and discrepant protein profiles in routinely applied laboratory tests.

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“…The most common method in proteomic analysis is two-dimensional gel electrophoresis coupled with mass spectrometry. Agarose gel electrophoresis (AGE) is the most commonly used method for clinical analysis of serum proteins [18]. Capillary electrophoresis (CE) is a more efficient technique and provides identification of a number of abnormalities which are not detected by AGE.…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoretic separation of serum proteins of workers occupationally exposed to heavy metals

Zalewska

Gawin

Ściskalska

et al. 2014

International Journal of Environmental Analytical Chemistry

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Metallurgy processes are associated with many hazardous and toxic factors, including heavy metals. Exposure to heavy metals can cause damage to different organs, which can be observed through variation in the concentration of proteins in serum. The aim of this study was to evaluate the changes in a serum protein profile of copper smelters exposed to As, Cd and Pb ions, and xenobiotics present in tobacco smoke. A 2.3-fold higher Pb concentration in the blood and a 2.8-fold greater As concentration have been observed in the urine of nonsmoking smelters compared to a control group. In the blood of smoking smelters, Cd concentration was 2-fold higher than in non-smoking ones. Serum proteins were separated by capillary electrophoresis, and in the group of non-smoking smelters, a higher amount of α 1 -globulins was observed. In the group of smoking smelters, fewer α 1 -globulins were noted. Furthermore, a greater amount of α 2 -globulins in the serum of smoking and non-smoking workers in relation to the control group was revealed. A positive correlation between the concentration of Cd in the blood and the content of a fraction containing α 1 -and α 2 -globulins was revealed. Urine Cd concentration was found to be negatively associated with the α 1 -and α 2 -globulins fraction. Observed abnormalities in the proteins profiles of smelters can be important markers when assessing exposure to heavy metals and in the early diagnosis of diseases caused by them.

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Performance Comparison of Capillary and Agarose Gel Electrophoresis for the Identification and Characterization of Monoclonal Immunoglobulins

Cited by 54 publications

References 11 publications

Comparison of the first fully automated agarose gel electrophoresis system with the Capillarys electrophoresis method for the identification of monoclonal immunoglobulins

Comparison of the first fully automated agarose gel electrophoresis system with the Capillarys electrophoresis method for the identification of monoclonal immunoglobulins

Gamma heavy chain disease in a patient with rheumatoid arthritis – a laboratory evaluation

Capillary electrophoretic separation of serum proteins of workers occupationally exposed to heavy metals

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