Magnetic particle-based chemiluminescence enzyme immunoassay for free thyroxine in human serum

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 50; no. 5; pp. 891 - 896
• Main Authors: Jin, Hui, Lin, Jin-Ming, Wang, Xu, Xin, Tian-bing, Liang, Shu-xuan, Li, Zhen-jia, Hu, Guo-mao
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 05.12.2009; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Binding, Competitive; Biological and medical sciences; Calibration; Chemiluminescence enzyme immunoassay; Fluorescein-5-isothiocyanate - pharmacology; Free thyroxine; Fundamental and applied biological sciences. Psychology; General pharmacology; Horseradish peroxidase; Horseradish Peroxidase - metabolism; Human serum; Humans; Hyperthyroidism - diagnosis; Hypothyroidism - diagnosis; Immunoenzyme Techniques - methods; Luminescence; Luminescent Measurements - methods; Magnetic particles; Magnetics; Medical sciences; Particle Size; Pharmacology. Drug treatments; Radioimmunoassay - methods; Reproducibility of Results; Thyroxine - blood; Horseradish peroxidase; Human serum; Magnetic particles; Free thyroxine; Chemiluminescence enzyme immunoassay; Human; Biological fluid; Enzyme; Aminoacid derivative hormone; Chemiluminescence; Enzyme immunoassay; Peroxidases; Thyroxine; Thyroid hormone; Peroxidase; Oxidoreductases
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2009.06.011

A magnetic particles-based chemiluminescence enzyme immunoassay with high sensitivity, specificity, rapidity, and reproducibility was developed for the determination of free thyroxine in human serum. A competitive assay has been proposed with horseradish peroxidase labeled thyroxine analog. The immunomagnetic particles coated with anti-fluorescein isothiocyanate antibody was used as dispersed solid phase and separation means for the immunoassay. Experimental conditions, such as temperature, the volume of magnetic particles and substrate, incubation time, dilution ratio and other relevant variables upon the immunoassay have been examined and optimized. The proposed method exhibited high performance which the linear range was 1.59–122 pmol L −1 and the detection limit was 0.25 pmol L −1. A coefficient of variance of less than 15% was obtained for both intra-assay and inter-assay precision. The present method has been successfully applied to the analysis of free thyroxine in human serum. The diagnostic accordance rate of the method for normal serum, hyperthyroidism and hypothyroidism are satisfactory. Good correlations were obtained between the results by the proposed method and the commercial radioimmunoassay kit. The present method exhibits good potential in the fabrication of FT 4 diagnostic kits which could be used in the clinical analysis and facilitated the development of automated operation systems in the clinical practice.