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 ◽  
...  

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.

Author(s):  
M A Jenkins ◽  
M D Guerin

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.


1998 ◽  
Vol 44 (6) ◽  
pp. 1184-1190 ◽  
Author(s):  
Yvonne Henskens ◽  
John de Winter ◽  
Maurits Pekelharing ◽  
Gabrielle Ponjee

Abstract Capillary electrophoresis (CE) and immunosubtraction capillary electrophoresis (IS-CE) were compared with the conventional methods agarose gel electrophoresis (AGE) and immunofixation electrophoresis (IFE) for detection and identification of paraproteins. In total, 74 paraproteins out of 468 serum samples were detected by both methods. Seventy-three monoclonal bands with concentrations ranging from 0.6 to 50.9 g/L were detected by the routine method. With CE, 70 paraproteins were detected and quantified on the electropherogram. Four paraproteins were not detected by CE; three of these were IgG (0.6, 1.1, and 2.2 g/L, respectively), and one was a IgM paraprotein (20.3 g/L) that could be visualized by minor changes in the running conditions. In comparison with IFE, 69 paraproteins were typed identically using IS-CE; only one paraprotein (IgMκ, 14.9 g/L) could not be identified. On the other hand, a monoclonal IgA band that had not been detected by AGE was identified by CE and IS-CE. We conclude that, in general, CE could be a useful method for detection of paraproteins and that IS-CE is a good alternative to IFE. Additional studies are required to investigate the ionic strength and pH of the running buffer, because these prove to be the most crucial factors for routine CE separation of paraproteins.


2018 ◽  
Vol 6 (2) ◽  
pp. 50-55
Author(s):  
Judith Gwladys Ekwe Priso ◽  
Jean Pierre Nda Mefo’o ◽  
Cécile Okalla Ebongue ◽  
Eveline Ngouadjeu Dongho Tsakeu ◽  
Catherine Akono Ndi ◽  
...  

Background: Electrophoresis of serum proteins is an orientation examination routinely used in clinical practice. For a few years, agarose gel electrophoresis has tended to be replaced with capillary electrophoresis owing to an increase in the accuracy of results. However, this technique is uncommon and is not widely used in Cameroon. Objectives: The research aimed at studying the electrophoretic profile of serum proteins using capillary technique among patients attending the Douala General Hospital, Cameroon. Methods: Capillary electrophoresis was used to carry out tests on blood samples from any inpatients and outpatients and fasting for 8-12 hours. Capillary electrophoresis of serum samples was used for the separation of proteins into six fractions and the total protidemia of each serum samples was determined using the Biuret method. Results were interpreted by observing the shape of curves and quantitative variations in each fraction of the different serum proteins. Results: A total of 311 patients participated in the study. The sampled population aged 50±18 years on average and consisted of 55.3% men and 44.7% women. All capillary electrophoresis profiles presented six protein fractions, namely, albumin, alpha (1 and 2), beta (1 and 2) and gamma globulins. Pathological disorders were diagnosed in 290 patients and 21 patients had normal results. Inflammatory syndromes accounted for 63.34% and monoclonal gammopathies for 10.29% the main pathological disorder identified. Conclusion: Capillary electrophoresis provides a more precise identification of biological syndromes and clear distinction of the six fractions of each protein. Monoclonal profiles and inflammatory syndromes were well detected. A prevalence of 10.29% was determined for gammopathies.


1998 ◽  
Vol 44 (6) ◽  
pp. 1191-1197 ◽  
Author(s):  
Thierry Le Bricon ◽  
Danielle Erlich ◽  
Djaouida Bengoufa ◽  
Michelle Dussaucy ◽  
Jean-Pierre Garnier ◽  
...  

Abstract We evaluated a new sodium dodecyl sulfate-agarose gel electrophoresis (SDS-AGE) for urinary protein analysis in patients with multiple myeloma (MM; n = 47; ages, 62 ± 2 years, mean ± SE). Abnormal proteinuria (mean = 1872 ± 360 mg/24 h) was present in 95% of the samples; 75% of the patients had some sign of renal dysfunction (glomerular and/or tubular) according to their SDS-AGE pattern. A band suggesting Bence Jones proteinuria (BJP) was detected in 40 vs 33 specimens by routine AGE. Immunofixation identified BJP in 38 patients; the calculated sensitivity of SDS-AGE for BJP was 97%. Excellent correlation (P <0.0001) was obtained with routine AGE (r = 0.994) and immunonephelometry (r = 0.963) for light chain quantification. SDS-AGE allows easy evaluation of renal dysfunction and shows high sensitivity for BJP detection. In a specialized laboratory, it is useful for following the progress of MM patients through the semiquantification of BJP.


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