A novel approach for a non competitive capillary electrophoresis immunoassay with laser-induced fluorescence detection for the determination of human serum albumin.
A new non competitive capillary electrophoresis immunoassay format based on a separation into a capillary modified by analyte immobilisation is described. The injection of an excess of labelled antibody off-line preincubated with the analyte allows the surface capture of the free antibody and the immunocomplex detection. It was developed using the human serum albumin (HSA) as analyte, two FITC-labelled antibodies and a HSA covalently linked capillary. Two calibration curves with good run-to-run reproducibility and LOD--respectively 14.0 nM for the FITC-polyclonal antibody and 9.0 nM for the FITC-monoclonal antibody--were achieved. The assay was applied to HSA determination in spiked samples of human urine with acceptable recoveries.
Giovannoli
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C
Department of Analytical Chemistry, University of Torino, Torino, Italy. cristina.giovannoli@unito.it <cristina.giovannoli@unito.it>
Anfossi
L
L
Baggiani
C
C
Giraudi
G
G
D000906:Antibodies / Q000276:immunology; D000936:Antigen-Antibody Complex; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D012709:Serum Albumin / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence
D000906:Antibodies; D000936:Antigen-Antibody Complex; D005456:Fluorescent Dyes; D012709:Serum Albumin