title: Microchip fluorescence-enhanced immunoaasay for simultaneous quantification of multiple tumor markers.
abstract: A microchip fluorescence-enhanced immunoassay method was developed for simultaneous detection of carcinoma antigen 125 (CA125) and carbohydrate antigen 15-3 (CA15-3). In this method, CA125 and CA15-3 react with excess amount of fluorescein isothiocyanate (FITC)-labeled monoclonal antibodies (Ab(*)) of CA125 and CA15-3 to form CA125-Ab(125)(*) and CA15-3-Ab(15-3)(*) complexes. Microchip electrophoresis (MCE) separation of free Ab(125)(*), Ab(15-3)(*), and CA125-Ab(125)(*), CA15-3-Ab(15-3)(*) complexes were then performed. The separated species were sensitively detected by laser-induced fluorescence detection (LIF). CA125 and CA15-3 were quantified simultaneously by measuring the fluorescence intensity of CA125-Ab(125)(*) and CA15-3-Ab(15-3)(*) complexes, respectively. Under the optimum conditions, the limits of detection were 0.23 U/mL for CA125 and 0.09 U/mL for CA15-3. The present MCE-LIF method was applied to the determination of CA125 and CA15-3 in serum from healthy subjects and cancer patients. The levels of CA125 and CA15-3 in these sera samples were found to be in the ranges of 15.6-36.1 U/mL and 13.8-28.4 U/mL for healthy subjects, and 192.5-368.3 U/mL and 63.3-198.4 U/mL for cancer patients.
authors.affiliation: Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 541004, China. zhaoshulin001@163.com
authors.forename: Ming
authors.initials: M
authors.lastname: Shi
authors.forename: Shulin
authors.initials: S
authors.lastname: Zhao
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Yi-Ming
authors.initials: YM
authors.lastname: Liu
authors.forename: Fanggui
authors.initials: F
authors.lastname: Ye
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D000936:Antigen-Antibody Complex / Q000737:chemistry; D014408:Biomarkers, Tumor / Q000097:blood*; D018394:CA-125 Antigen / Q000097:blood; D046209:Electrophoresis, Microchip / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D018396:Mucin-1 / Q000097:blood; D009369:Neoplasms / Q000175:diagnosis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D014408:Biomarkers, Tumor; D018394:CA-125 Antigen; D018396:Mucin-1; D012967:Sodium Dodecyl Sulfate; D016650:Fluorescein-5-isothiocyanate