| 摘 要： | A novel, direct noncompetitive flow injection enzyme immunoassay for α-fetoprotein (AFP) was developed by enhanced chemiluminescence
detection. The method was based on off-line incubation of AFP and horseradish peroxidase (HRP)-labeled anti-AFP, and then
trapping of the unbound enzyme conjugate by an immunoaffinity column filled with AFP-modified Sepharose. The immunocomplex
formed in incubation passed through the column and then was directly detected by a postcolumn chemiluminescence technique.
The optimal conditions for the immunoassay procedure and chemiluminescence detection were established. At a 1:10 dilution
of enzyme conjugate solution, the linear range for chemiluminescence detection of AFP was from 2.0 to 75 ng/mL with a correlation
coefficient of 0.993 and a coefficient of variation of 2.67% at 30 ng/mL. The detection limit was 0.5 ng/mL. This method was
flexible, sensitive, and rapid. The immunoaffinity column of 200 μL could be repeatedly used 100 times without a single decrease.
The whole assay time including the preincubation step was only 30 min for one sample.
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