A fluorescence polarization immunoassay (FPIA) was developed for the analysis ofaflatoxins (AFs) using an anti-aflatoxin B1 (AFB1) monoclonal antibody and a novel fluorescein-labeled AFB1 tracer. The FPIA showed...A fluorescence polarization immunoassay (FPIA) was developed for the analysis ofaflatoxins (AFs) using an anti-aflatoxin B1 (AFB1) monoclonal antibody and a novel fluorescein-labeled AFB1 tracer. The FPIA showed an IC50 value of 23.33 ng/mL with a limit of detection of 13.12 ng/mL for AFB1. The cross-reactivities of AFB1, AFB2, AFG1, AFG2, AFM1, and AFM2 with the antibody were 100%, 65.7%, 143%, 23.5%, 111.4%, and 2%, respectively. The group-specificity of anti-AFB1mAb indicated that the FPIA could potentially be used in a screening method for the detection of total AFs, albeit not AFG2 and AFM2. The total time required for analyzing 96 samples in one microplate was less than 5 rain. This study demonstrates the potential usefulness of the FPIA as a rapid and simple technique for monitoring AFs.展开更多
Estrone has been identified as a potential endocrine-disrupting chemical (EDC)[1]. Estrone is usually quantified by gas chromatography-mass spectrometry (GC-MS), GC-MS/MS, high performance liquid chromatography (...Estrone has been identified as a potential endocrine-disrupting chemical (EDC)[1]. Estrone is usually quantified by gas chromatography-mass spectrometry (GC-MS), GC-MS/MS, high performance liquid chromatography (HPLC), HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay (ELISA) or chemiluminescence immunoassay (CLIA) for determination of estrone in real samples have been developed[2'4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods (with separation) to homogeneous assays (without separation)[5]. Fluorescence polarization immunoassay (FPIA) is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production.展开更多
A fluorescence polarization immunoassay(FPIA) for the determination of salinomycin(SAL) was developed by using anti-SAL monoclonal antibodies(mAb).Fluorescein labeled SAL(tracer) was synthesized by the N-hydroxysuccin...A fluorescence polarization immunoassay(FPIA) for the determination of salinomycin(SAL) was developed by using anti-SAL monoclonal antibodies(mAb).Fluorescein labeled SAL(tracer) was synthesized by the N-hydroxysuccinimide active ester method and purified using thin layer chromatography(TLC).The developed FPIA for SAL had a dynamic range from 0.60 to 2193 ng/mL with an IC50 value of 33.2 ng/mL and a detection limit(LOD) of 0.08 ng/mL.No significant cross-reactivities were observed with other drugs but 67.6% with narasin.展开更多
Objective: This study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum. Methods: To measure the concentration of theophylline (n=122) and eval...Objective: This study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum. Methods: To measure the concentration of theophylline (n=122) and evaluate the assay.Results: The linear range of the CLIA method was 0.51~40 mg/L (Y=1.02X+0.44, r=0.995). The intra and inter CV (coefficient variance) of CLIA were 3.20% and 3.57%, respectively. The average recovery rate was 102.3%. This method was free from interference by brilirubin (<200 μmol/L), hemoglobin (<10 g/L), and triglycerides (<15 mmol/L). Conclusion: This method is simple, convenient and precise for clinical pharmacokinetics study oftheophylline.展开更多
基金supported by grants from the International Science&Technology Cooperation Program of China(2009DFA32330)the Special Fund for Agro-scientific Research in the Public Interest(No.201203040)
文摘A fluorescence polarization immunoassay (FPIA) was developed for the analysis ofaflatoxins (AFs) using an anti-aflatoxin B1 (AFB1) monoclonal antibody and a novel fluorescein-labeled AFB1 tracer. The FPIA showed an IC50 value of 23.33 ng/mL with a limit of detection of 13.12 ng/mL for AFB1. The cross-reactivities of AFB1, AFB2, AFG1, AFG2, AFM1, and AFM2 with the antibody were 100%, 65.7%, 143%, 23.5%, 111.4%, and 2%, respectively. The group-specificity of anti-AFB1mAb indicated that the FPIA could potentially be used in a screening method for the detection of total AFs, albeit not AFG2 and AFM2. The total time required for analyzing 96 samples in one microplate was less than 5 rain. This study demonstrates the potential usefulness of the FPIA as a rapid and simple technique for monitoring AFs.
基金supported by Natural Science Foundation of China(U1301214)Guangdong Natural Science Foundation(S2013030013338)+1 种基金the PhD Programs Foundation of Ministry of Education of China(20114404130002)National Department Public Benefit Research Foundation(201003008-08)
文摘Estrone has been identified as a potential endocrine-disrupting chemical (EDC)[1]. Estrone is usually quantified by gas chromatography-mass spectrometry (GC-MS), GC-MS/MS, high performance liquid chromatography (HPLC), HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay (ELISA) or chemiluminescence immunoassay (CLIA) for determination of estrone in real samples have been developed[2'4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods (with separation) to homogeneous assays (without separation)[5]. Fluorescence polarization immunoassay (FPIA) is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production.
基金support from the National Natural Science Foundation of China (Grant No 30830082)the Major State Basic Research Development Program of China (Grant No 2009CB118801)Beijing Excellent Doctoral Dissertation Fund (Grant No YB20081001902)
文摘A fluorescence polarization immunoassay(FPIA) for the determination of salinomycin(SAL) was developed by using anti-SAL monoclonal antibodies(mAb).Fluorescein labeled SAL(tracer) was synthesized by the N-hydroxysuccinimide active ester method and purified using thin layer chromatography(TLC).The developed FPIA for SAL had a dynamic range from 0.60 to 2193 ng/mL with an IC50 value of 33.2 ng/mL and a detection limit(LOD) of 0.08 ng/mL.No significant cross-reactivities were observed with other drugs but 67.6% with narasin.
文摘Objective: This study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum. Methods: To measure the concentration of theophylline (n=122) and evaluate the assay.Results: The linear range of the CLIA method was 0.51~40 mg/L (Y=1.02X+0.44, r=0.995). The intra and inter CV (coefficient variance) of CLIA were 3.20% and 3.57%, respectively. The average recovery rate was 102.3%. This method was free from interference by brilirubin (<200 μmol/L), hemoglobin (<10 g/L), and triglycerides (<15 mmol/L). Conclusion: This method is simple, convenient and precise for clinical pharmacokinetics study oftheophylline.