scholarly journals Analysis of serum proteins by agarose gel and capillary electro phoresis

2013 ◽  
Vol 26 (3) ◽  
pp. 267-272
Keyword(s):  
Capillary Electrophoresis ◽  
Serum Protein ◽  
Gel Electrophoresis ◽  
Protein Fraction ◽  
High Speed ◽  
Serum Proteins ◽  
Attractive Alternative ◽  
Automatic Process ◽  
Agarose Gel ◽  
Two Systems

Electrophoresis is a basic technique used to identify disorders of blood serum protein fractions. Agarose gel is the most frequently used medium for routine protein separations. However, capillary electrophoresis seems to be an attractive alternative to gel electrophoresis. The article presents the results of comparative analysis of two systems (Sebia): Hydrasys designed for electrophoretic separations on agarose gel and Minicap for capillary electrophoresis. The purpose of study was to evaluate comparatively the concentrations of each serum protein fraction obtained by gel and capillary electrophoresis and to analyze the correlations between the results obtained by those two systems depending on the concentrations of each protein fraction. The study was carried out in the group of 98 patients, 46 females and 52 males. Despite slight quantitative differences in certain fractions obtained by both methods, capillary electrophoresis offers a fully automatic process of analysis, high speed and efficiency which proves that capillary electrophoresis is appropriate alternative to gel electrophoresis.

Quantitative analysis of serum proteins separated by capillary electrophoresis

1993 ◽  
Vol 39 (4) ◽  
pp. 689-692 ◽  
Author(s):  
J W Kim ◽  
J H Park ◽  
J W Park ◽  
H J Doh ◽  
G S Heo ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Serum Samples ◽  
Polyclonal Gammopathy ◽  
Quantitative Analyses

Abstract The possibility of open tubular capillary electrophoresis for clinical diagnostic use is examined. Capillary electrophoresis was performed in an untreated 50 microns (i.d.) x 100 cm (65 cm to detector) capillary with detection of absorbance at 200 nm. Conditions for the separation of serum proteins without adsorption to the capillary surface were established. Quantitative analyses of serum samples from 38 patients with liver cirrhosis, nephrotic syndrome, or polyclonal gammopathy by capillary electrophoresis were done and the results were compared with those by conventional agarose gel electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All samples were analyzed in duplicate. We evaluated linearity of response, within-run CV, and the correlation between capillary electrophoresis and agarose gel electrophoresis.

scholarly journals Dual Detection High-Speed Capillary Electrophoresis for Simultaneous Serum Protein Analysis and Immunoassays

2021 ◽  
Author(s):  
Prabhavie Opallage ◽  
Miyuru De Silva ◽  
Robert Dunn
Keyword(s):  
Refractive Index ◽  
Capillary Electrophoresis ◽  
Human Serum ◽  
Serum Protein ◽  
High Speed ◽  
Serum Proteins ◽  
Numerical Aperture ◽  
Excitation Source ◽  
Detection Zone ◽  
Dual Detection

Abstract Serum protein electrophoresis (SPE) and immunoassays are important tools used clinically to diagnose disease. SPE separates serum proteins into bands whose shape and amplitude can alert clinicians to a range of disorders. This is usually followed by more specific immunoassays to quantify important antigens and confirm a diagnosis. Here we develop a high-speed capillary electrophoresis (HSCE) platform capable of both SPE measurements and quantifying immunoassays, simultaneously. The HSCE uses a 10 cm long total length separation capillary (50 μm i.d., 80 μm o.d.) with an 8 cm length-to-detector for rapid analysis times. This is important for throughput in clinical settings and for quantifying immunoassays, where antigen-antibody complexes continually dissociate once injected into the non-equilibrium conditions of the separation capillary. A single laser excitation source is focused into the detection zone of the capillary to measure both refractive index (SPE) and fluorescence signals (immunoassays), simultaneously. Light scattered back towards the excitation source measures refractive index changes using back-scatter interferometry (BSI), while fluorescence is collected from below the capillary with a high numerical aperture objective. To validate the dual detection HSCE approach, SPE and immunoassays are measured from human serum samples pre-incubated with fluorescein and an anti-fluorescein monoclonal antibody. We show that the BSI signal measures characteristic SPE profiles for human serum separated in 100 mM boric acid (pH 10), 100 mM arginine (pH 11), and 20 mM CHES (pH 10). For the immunoassay, the fluorescence electropherograms reveal that CHES provides the optimal buffer for measuring the immunocomplex and separating it from the free antigen. Immunoassays in CHES yield a LOD of 23 nM and a LOQ of 70 nM for the detection of fluorescein. Elevated pH is used in SPE to ensure all proteins are charged and to reduce protein adsorption to the capillary walls. The high pH, however, also reduces antibody affinity. Preliminary studies carried out in 50 mM barbital at pH 8 show improved stability of the immunocomplex and better separation for immunoassay quantification, but loss of resolution in the SPE. Further optimization will open new capabilities for measuring orthogonal diagnostic signals in seconds with HSCE.

scholarly journals Quantification of Serum Proteins Using Capillary Electrophoresis

1995 ◽  
Vol 32 (5) ◽  
pp. 493-497 ◽  
Author(s):  
M A Jenkins ◽  
M D Guerin
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Clinical Laboratory ◽  
Charged Particles ◽  
Protein Electrophoresis ◽  
Agarose Gel Electrophoresis ◽  
Serum Samples ◽  
Routine Clinical Laboratory ◽  
Sample Dilution

Capillary electrophoresis is a technique that can be automated for the separation of charged particles. By investigating suitable sample dilution and injection time and adhering to a strict washing procedure we have been able to quantify paraproteins in serum samples. This has enabled us to use the technique of capillary electrophoresis for the provision of serum protein electrophoresis in a routine clinical laboratory. We present our findings of 260 serum samples, which included 76 samples with paraproteins analysed by both capillary electrophoresis (EC) and high resolution agarose gel electrophoresis (HRAGE). CE was able to detect all the monoclonal bands detected by HRAGE, and, in particular, better able to detect IgA monoclonal bands occurring in the beta region. The major advantages of CE over HRAGE relate to the automated nature of CE with the elimination of the need for a densitometer.

Absence of beta-globulin band in the serum protein electropherogram of a patient with liver disease.

1981 ◽  
Vol 27 (2) ◽  
pp. 334-336 ◽  
Author(s):  
A O Vladutiu ◽  
J S Kim
Keyword(s):  
Liver Cirrhosis ◽  
Liver Disease ◽  
Serum Protein ◽  
Gel Electrophoresis ◽  
Marked Decrease ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Glycoprotein I ◽  
Complement Component C3 ◽  
Beta Globulin

Abstract Agarose-gel electrophoresis of serum of a 72-year-old woman with liver cirrhosis showed virtually no beta-globulins two weeks before the patient's death. There was marked decrease in the concentrations of transferrin, beta-lipoproteins, hemopexin, complement component C3, beta-glycoprotein I, and cholesterol in serum. Absence of a beta-globulin band appears to signify an ominous prognosis.

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