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A sensitive immunoassay for determination of hepatitis B surface antigen and antibody in human serum using capillary electrophoresis with chemiluminescence detection.
- Source :
-
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2007 Sep 15; Vol. 857 (1), pp. 100-7. Date of Electronic Publication: 2007 Jul 10. - Publication Year :
- 2007
-
Abstract
- A sensitive and homogeneous immunoassay (IA) based on capillary electrophoresis (CE) with enhanced chemiluminescence (CL) detection has been developed for the determination of hepatitis B surface antigen (HBsAg) and antibody (HBsAb) in human serum. The conditions for the CL reaction and electrophoresis were investigated in detail using horseradish peroxidase (HRP) labeled HBsAg (HBsAg*) as a marker because of its catalytic effects on the luminol-hydrogen peroxide reaction. The CL reaction was enhanced by para-iodophenol and the CL detector was designed uniquely without any dead volume or diluents effect. The present method has been used for assaying HBsAg and HBsAb in human serum using a competitive format and a non-competitive format, respectively. Under the optimal conditions, the linear ranges were from 1 to 400 pmol/L (R=0.9988) for HBsAg and 2 to 200 mIU/mL (R=0.9981) for HBsAb. The detection limits were 0.4 pmol/L and 1 mIU/mL for HBsAg and HBsAb, respectively. The relative standard deviations of peak area were 4.2% and the errors of it were from -0.03% to +0.05% for 80 pmol/L HBsAg* (n=7). In this study, the free HBsAg* and the bound HBsAg* (HBsAg*-HBsAb) were separated in the separation capillary within 6 min using a borate run buffer. To verify the experimental reliability, the result was comparable with that of enzyme linked immunosorbent assay (ELISA) and demonstrated the feasibility of the CE-CL immunoassay method for clinical diagnosis.
- Subjects :
- Antigen-Antibody Complex chemistry
Binding, Competitive
Feasibility Studies
Hepatitis B diagnosis
Horseradish Peroxidase metabolism
Humans
Hydrogen Peroxide chemistry
Immunoenzyme Techniques
Iodobenzenes chemistry
Luminescence
Luminescent Measurements instrumentation
Luminol chemistry
Reference Standards
Sensitivity and Specificity
Electrophoresis, Capillary methods
Hepatitis B Antibodies blood
Hepatitis B Surface Antigens blood
Luminescent Measurements methods
Subjects
Details
- Language :
- English
- ISSN :
- 1570-0232
- Volume :
- 857
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
- Publication Type :
- Academic Journal
- Accession number :
- 17689153
- Full Text :
- https://doi.org/10.1016/j.jchromb.2007.07.006