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Clinical Chemistry 49: 1909-1915, 2003; 10.1373/clinchem.2003.017756
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(Clinical Chemistry. 2003;49:1909-1915.)
© 2003 American Association for Clinical Chemistry, Inc.

Automation and Analytical Techniques

Automated Multicapillary Electrophoresis for Analysis of Human Serum Proteins

Cécile Gay-Bellile1, Djaouida Bengoufa2, Pascal Houze1, Didier Le Carrer3, Mourad Benlakehal1, Bernard Bousquet1, Bernard Gourmel1 and Thierry Le Bricon1,a

1 Laboratoire de Biochimie A et de Neurobiologie and
2 Laboratoire d’Immunologie et d’Histocompatibilité, Hôpital St-Louis AP-HP, 1 avenue Claude Vellefaux, 75010 Paris, France.

3 Laboratoire de Biochimie Spécialisée, Hotel Dieu, 9 quai Moncousu, 44093 Nantes Cedex 1, France.

aAuthor for correspondence. Fax 33-1-4249-9247; e-mail thierry.le-bricon{at}sls.ap-hop-paris.fr.

Background: We evaluated a new, automated multicapillary zone electrophoresis (CE) instrument (Capillarys®, 4.51 software version; Sebia) for human serum protein analysis.

Methods: With the Capillarys ß1-ß2+® reagent set, proteins were separated at 7 kV for 4 min in 15.5 cm x 25 µm fused-silica capillaries (n = 8) at 35.5 °C in a pH 10 buffer with online detection at 200 nm. Serum samples with different electrophoretic patterns (n = 265) or potential interference (n = 69) were analyzed and compared with agarose gel electrophoresis (AGE; Hydrasys®-Hyrys®, Hydragel protein(e) 15/30® reagent set; Sebia).

Results: CVs were <3.5% for albumin, <11% for {alpha}1-globulin, <4.1% for {alpha}2-globulin, <7.4% for ß-globulin, and <5.8% for {gamma}-globulin (3 control levels); measured throughput was 60 samples/h. In patients without paraprotein (n = 116), the median differences between CE and AGE were -5.4 g/L for albumin, 4.0 g/L for {alpha}1-globulin, 0.7 g/L for {alpha}2-globulin, 0.6 g/L for ß-globulin (P <0.001 for all fractions), and -0.1 g/L for {gamma}-globulin (not significant). More samples had at least one {gamma}-migrating peak detected by CE (n = 135 vs 130; paraprotein detection limit, ~0.5–0.7 g/L), but fewer were quantified (n = 84 vs 91) because of {gamma}- to ß-migration shifts. There was a 1.2 g/L median difference between CE and AGE for {gamma}-migrating paraprotein quantification (n = 69; P <0.001). Several ultraviolet-absorbing substances (lipid emulsion, hemoglobin) or molecules (contrast agent, gelatin-based plasma substitute) induced CE artifacts.

Conclusions:The Capillarys instrument is a reliable CE system for serum protein analysis, combining advantages of full automation (ease of use, bar-code identification, computer-assisted correction of {alpha}1-globulins) with high analytical performances and throughput.




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