BACKGROUND: Human serum albumin (HSA) is the most abundant plasma protein and plays key a role in metabolism. The variation in albumin concentration provides valuable information related to metabolic diseases and diagnostic application. METHODS: We constructed two assay systems to quantify the albumin concentration. The immunoassay used a fluorescence (FL) dye to detect albumin in samples and employed the conventional chromatography as a separation system. The assay system consists of an anti-HSA-mAb or an HSA immobilized test strip in a disposable cartridge, a fluorescence-labeled detector buffer and a laser-fluorescence scanner. We mixed the sample with detector, loaded it onto a cartridge, incubated it for 10 min and measured the concentration of albumin in a laser-fluorescence scanner. We examined the comparability of assay with an automated BCG dye binding method using a Hitachi 747 biochemical analyzer. RESULTS: The correlation of coefficient between AT/AC as converted from the relative fluorescence units (RFU) and albumin concentration displayed reasonable reliability in both the competition and the inhibition assay systems (r = 0.998). Using the Bland-Altman difference plot analysis, we observed an acceptable agreement between two methods, the fluorescence immunochromatography assay (FL-ICA) and the automated BCG dye-binding method of a Hitachi biochemical analyzer, over the clinical relevant range of HSA concentrations. The coefficient of variation (CV) of within- and between-run variation in the immunoassay system was < 8% and the recovery fell within 5% in each control sample. In addition to its reliable analytical performance, the assay with whole blood can be completed in 12 min using a one-step operation without any pretreatment. CONCLUSION: The developed immunoassay system using fluorescence dye and lateral-flow chromatography is a simple, fast and reliable method for quantifying the albumin concentration in whole blood.
BACKGROUND:Humanserum albumin (HSA) is the most abundant plasma protein and plays key a role in metabolism. The variation in albumin concentration provides valuable information related to metabolic diseases and diagnostic application. METHODS: We constructed two assay systems to quantify the albumin concentration. The immunoassay used a fluorescence (FL) dye to detect albumin in samples and employed the conventional chromatography as a separation system. The assay system consists of an anti-HSA-mAb or an HSA immobilized test strip in a disposable cartridge, a fluorescence-labeled detector buffer and a laser-fluorescence scanner. We mixed the sample with detector, loaded it onto a cartridge, incubated it for 10 min and measured the concentration of albumin in a laser-fluorescence scanner. We examined the comparability of assay with an automated BCG dye binding method using a Hitachi 747 biochemical analyzer. RESULTS: The correlation of coefficient between AT/AC as converted from the relative fluorescence units (RFU) and albumin concentration displayed reasonable reliability in both the competition and the inhibition assay systems (r = 0.998). Using the Bland-Altman difference plot analysis, we observed an acceptable agreement between two methods, the fluorescence immunochromatography assay (FL-ICA) and the automated BCG dye-binding method of a Hitachi biochemical analyzer, over the clinical relevant range of HSA concentrations. The coefficient of variation (CV) of within- and between-run variation in the immunoassay system was < 8% and the recovery fell within 5% in each control sample. In addition to its reliable analytical performance, the assay with whole blood can be completed in 12 min using a one-step operation without any pretreatment. CONCLUSION: The developed immunoassay system using fluorescence dye and lateral-flow chromatography is a simple, fast and reliable method for quantifying the albumin concentration in whole blood.
Authors: Evgenia G Matveeva; Ignacy Gryczynski; Joanna Malicka; Zygmunt Gryczynski; Ewa Goldys; Joseph Howe; Klaus W Berndt; Joseph R Lakowicz Journal: J Fluoresc Date: 2005-11 Impact factor: 2.217
Authors: Evgenia G Matveeva; Zygmunt Gryczynski; Joanna Malicka; Joanna Lukomska; Slawomir Makowiec; Klaus W Berndt; Joseph R Lakowicz; Ignacy Gryczynski Journal: Anal Biochem Date: 2005-09-15 Impact factor: 3.365
Authors: Bin Liu; Xiaoman Bi; Lucas McDonald; Yi Pang; Danqing Liu; Chengjun Pan; Lei Wang Journal: Sens Actuators B Chem Date: 2016-06-11 Impact factor: 7.460