Investigation of a capillary electrophoretic approach for direct quantification of apolipoprotein A-I in serum.
In the present study a rapid, reproducible and robust capillary electrophoresis (CE) procedure for the quantification of apolipoprotein A-I (Apo A-I) in serum without pretreatment has been developed (total run time, 11 min). The coefficients of variation (CV; n = 10) for the relative peak area are 1.8% at a concentration of 145 mg/dL and 1.6% at 196 mg/dL; and for the inter-assay 8.9% at 161 mg/dL (10 consecutive days), i.e., similar to the CVs of a high-throughput immunonephelometric routine assay. The CV for the migration time is 0.4% (n = 20). The robustness of the CE approach was tested in patient samples with hemolysis, hyperbilirubinemia and hyperlipidemia. A comparison of 99 Apo A-I serum values with results of a fixed-time immunonephelometric routine assay showed a positive constant bias of 60% (mean) for the immunonephelometric values, no deviation from linearity, but significant deviations in several samples. Investigations on interferences in the CE analyses gave no evidence that CE failed. Our study shows that CE is amenable to a fast analysis and a reproducible and reliable quantification of Apo A-I level in sera of various clinical samples.
Klinisch-chemisches Zentrallabor der Universitätskliniken, Abteilung Innere Medizin IV Universität Tübingen, Germany. rainer.lehmann@med.uni-tuebingen.de
Rainer
R
Lehmann
Ulrich
U
Friess
Hans-Ulrich
HU
Häring
Wolfgang
W
Voelter
Hartmut
H
Liebich
Alexander
A
Beck
D016632:Apolipoprotein A-I / Q000097:blood*; D002138:Calibration; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D006461:Hemolysis; D006801:Humans; D006932:Hyperbilirubinemia / Q000097:blood; D006949:Hyperlipidemias / Q000097:blood; D007118:Immunoassay; D015203:Reproducibility of Results; D013997:Time Factors
D016632:Apolipoprotein A-I