Abstract
A flow injection chemiluminescence immunoassay for rapid and sensitive detection of carcinoembryonic antigen (CEA) by using a phenylboronic acid-based immunoaffinity column as a glycoprotein collector was proposed in this paper. The column was prepared by coupling of 3-aminophenylboronic acid on the glass beads through a γ-glycidoxypropyltrimethoxysilane (GPMS) linkage. Based on an indirect competitive immunoreaction, the mixture of CEA sample and enzyme conjugated CEA antibody (HRP-anti-CEA) was incubated in advance, followed by direct injection to the column to capture free HRP-labeled CEA antibody in the column. The trapped HRP-labeled antibody was detected by flow inject chemiluminescence in the presence of luminol and hydrogen peroxide. The decreased chemiluminescent signal was proportional to the concentration of CEA in the range of 3.0–30.0 ng/mL with a correlation coefficient of 0.998. The column showed an acceptable reproducibility and stability and is potentially used for practical clinical detection of the serum CEA level.
Highlights
Carcinoembryonic antigen (CEA) is a highly glycosylated cell surface glycoprotein with a total of 28 asparagine-linked glycosylation sites [1]
The 3-aminophenylboronic acid (APBA) coated glass microbeads were prepared by using γ-glycidoxypropyltrimethoxysilane (GPMS) as linkage
The as-prepared APBA modified microbeads were filled into a glassy tube followed by injection of a 250 ng/mL carcinoembryonic antigen (CEA) solution into the column to form the immunoaffinity column
Summary
Carcinoembryonic antigen (CEA) is a highly glycosylated cell surface glycoprotein with a total of 28 asparagine-linked glycosylation sites [1]. Enzyme-linked immunoassays [3,4], fluorescence measurement [5,6,7] and chemiluminescence assay [8] are frequently used to detect to successfully detect the CEA in practical samples with high selectivity, sufficient sensitivity and precision. These conventional immunoassay methods need enzyme or fluorescent-labeled antibody/antigen, not to mention its lengthy analysis that requires highly skilled personnel, specially equipped laboratories, and expensive chemicals [9,10,11]. The trapped HRP-labeled CEA antibody enhanced the chemiluminescence emission intensity of luminal system, which is the basic for detection of the analyte CEA in samples
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