Automated multicapillary electrophoresis for analysis of human serum proteins.
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 beta1-beta2+ reagent set, proteins were separated at 7 kV for 4 min in 15.5 cm x 25 micro m fused-silica capillaries (n = 8) at 35.5 degrees 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 beta-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 beta-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, approximately 0.5-0.7 g/L), but fewer were quantified (n = 84 vs 91) because of gamma- to beta-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.
Gay-Bellile
Cécile
C
Laboratoire de Biochimie A et de Neurobiologie, Hôpital St-Louis AP-HP, 1 avenue Claude Vellefaux, 75010 Paris, France.
Bengoufa
Djaouida
D
Houze
Pascal
P
Le Carrer
Didier
D
Benlakehal
Mourad
M
Bousquet
Bernard
B
Gourmel
Bernard
B
Le Bricon
Thierry
T
D000510:Alpha-Globulins / Q000032:analysis; D001322:Autoanalysis; D001609:Beta-Globulins / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods; D006801:Humans; D010266:Paraproteins / Q000032:analysis; D012709:Serum Albumin / Q000032:analysis; D005719:gamma-Globulins / Q000032:analysis
D000510:Alpha-Globulins; D001609:Beta-Globulins; D001798:Blood Proteins; D010266:Paraproteins; D012709:Serum Albumin; D005719:gamma-Globulins