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Noncompetitive enzyme immunoassay for α-fetroprotein using flow injection chemiluminescence

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Abstract

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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Authors and Affiliations

  1. State Key Laboratory of Coordination Chemistry, Department of Chemistry, Institute of Chemical Biology, Nanjing University, 210093, Nanjing, P. R. China

    Jiehua Lin & Huangxian Ju

  2. Jiangsu Institute of Cancer Prevention and Cure, 210009, Nanjing, P. R. China

    Feng Yan

Authors
  1. Jiehua Lin
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  2. Feng Yan
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  3. Huangxian Ju
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Correspondence to Huangxian Ju.

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Lin, J., Yan, F. & Ju, H. Noncompetitive enzyme immunoassay for α-fetroprotein using flow injection chemiluminescence. Appl Biochem Biotechnol 117, 93–102 (2004). https://doi.org/10.1385/ABAB:117:2:093

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  • DOI: https://doi.org/10.1385/ABAB:117:2:093

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