ABSTRACT
A piezoelectric-based immunosensor was developed for high density lipoprotein particle (HDL-P) measurement. Monoclonal anti-human apolipoprotein A1 antibody was used as a specific binding molecule for the major apolipoprotein of HDL-P. This sensing element was fabricated by immobilizing the anti-human apolipoprotein A1 on a 12 MHz AT-cut quartz crystal via a 3-mercaptopropionic acid (MPA) self-assembled monolayer. The frequency shift from the mass change of the antigen-antibody binding refers to the amount of HDL-P. The optimal antibody immobilization was performed to achieve the maximum potential of the antibody. The appropriate quantity and immobilization time of the antibody were 0.1 mg ml(-1) and 90 minutes, respectively. The immobilized antibody in the HDL-P immunosensor accomplished perfect binding with HDL-P within 60 minutes. The dose-response curve for HDL-P showed a linear response from 0.21 to 2.50 mg protein per ml equivalent to 0.40 × 10(10) to 3.65 × 10(10) particles per µl without significant interference from other lipoproteins. The intra- and inter-assay imprecision (CV) were 7.8 and 18.5%, respectively. The analytical accuracy of this measurement was 96.29-96.31%. The HDL-P concentration obtained from the sensor revealed a 2.05 mg protein per ml with 0.26 mg protein per ml of expanded uncertainty at the 95% confidence level. This immunosensor gave an assay result which correlated with the homogeneous enzymatic colorimetric assay (R(2) = 0.902).
Subject(s)
Antibodies, Immobilized/chemistry , Electrochemical Techniques/instrumentation , Immunoassay/instrumentation , Lipoproteins, HDL/blood , 3-Mercaptopropionic Acid/chemistry , Apolipoprotein A-I/analysis , Apolipoprotein A-I/blood , Biosensing Techniques/instrumentation , Crystallization , Equipment Design , Humans , Limit of Detection , Lipoproteins, HDL/analysis , Quartz/chemistryABSTRACT
BACKGROUND: A centrifuge-based device has been introduced to the Samsung Blood Analyzer (SBA). The verification of this analyzer is essential to meet the ISO15189 standard. METHODS: Analytical performance was evaluated according to the NCCLS EP05-A method. The results of plasma samples were compared between the SBA and a Hitachi 917 analyzer according to the NCCLS EP09-A2-IR method. Percent recovery was determined via analysis of original control serum and spiked serum. RESULTS: Within-run precision was found to be 0.00 - 6.61% and 0.96 - 5.99% in normal- and abnormal-level assays, respectively, while between-run precision was 1.31 - 9.09% and 0.89 - 6.92%, respectively. The correlation coefficients (r) were > 0.990. The SBA presented analytical accuracy at 96.64 +/- 3.39% to 102.82 +/- 2.75% and 98.31 +/- 4.04% to 103.61 +/- 8.28% recovery, respectively. CONCLUSIONS: The results obtained verify that all of the 13 tests performed using the SBA demonstrates good and reliable precision suitable for use in qualified clinical chemistry laboratory service.
