An indirect perfluorosulfonated ionomer-coated electrochemical immunosensor for the detection of the protein human chorionic gonadotrophin

被引:59
作者
Chetcuti, AF
Wong, DKY [1 ]
Stuart, MC
机构
[1] Macquarie Univ, Sch Chem, Sydney, NSW 2109, Australia
[2] Macquarie Univ, Dept Biol Sci, Sydney, NSW 2109, Australia
关键词
D O I
10.1021/ac981216a
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The development of an amperometric immunosensor for the detection of human chorionic gonadotrophin (hCG) is described. Ln this immunosensor, Nafion was used to immobilize an anti-hCG monoclonal antibody onto a glassy carbon electrode. A systematic study on the effects of experimental parameters such as the quantity of ethanol present in the Nafion solution, the percentage composition of Nafion, the pH of the immobilization buffer, and the concentration of antibody used for entrapment experiments on the binding between the immobilized antibody and I-125-labeled hCG has been carried out. Two immobilization methods, coimmobilization and adsorption immobilization, have then been attempted. A binding of approximately 3% was obtained in the former method, while 5.5% binding was achieved in the latter. On the basis of these results, adsorption immobilization was employed to entrap antibody on the electrode surface. A sandwich assay was then developed for hCG in which the enzyme horseradish peroxidase was conjugated to a second anti-hCG monoclonal antibody. The activity of the enzyme was determined electrochemically by the reduction of benzoquinone to hydroquinone. Binding of hCG to immobilized antibody determines the quantity of enzyme-conjugated antibody at the electrode surface, permitting the quantification of hCG. By a standard additions calibration method of hCG performed in blank human serum samples, the immunosensor exhibits a limit of linearity at 200 mIU mL(-1) and a detection limit of 11.2 mIU mL(-1) (based on twice the standard deviation of the blank solution).
引用
收藏
页码:4088 / 4094
页数:7
相关论文
共 30 条
  • [1] Development of a new immunosensor for pesticide detection: a disposable system with liposome-enhancement and amperometric detection
    Baumner, AJ
    Schmid, RD
    [J]. BIOSENSORS & BIOELECTRONICS, 1998, 13 (05) : 519 - 529
  • [2] SPECIFICITY OF 2-SITE IMMUNOASSAYS
    BOSCATO, LM
    EGAN, GM
    STUART, MC
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 1989, 117 (02) : 221 - 229
  • [3] SERUM HUMAN CHORIONIC-GONADOTROPIN LEVELS THROUGHOUT NORMAL-PREGNANCY
    BRAUNSTEIN, GD
    RASOR, J
    ADLER, D
    DANZER, H
    WADE, ME
    [J]. AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY, 1976, 126 (06) : 678 - 681
  • [4] ECTOPIC PRODUCTION OF HUMAN CHORIONIC GONADOTROPIN BY NEOPLASMS
    BRAUNSTEIN, GD
    VAITUKAITIS, JL
    CARBONE, PP
    ROSS, GT
    [J]. ANNALS OF INTERNAL MEDICINE, 1973, 78 (01) : 39 - 45
  • [5] COLE LA, 1992, CLIN CHEM, V38, P263
  • [6] SEPARATION-FREE SANDWICH ENZYME IMMUNOASSAYS USING MICROPOROUS GOLD ELECTRODES AND SELF-ASSEMBLED MONOLAYER IMMOBILIZED CAPTURE ANTIBODIES
    DUAN, CM
    MEYERHOFF, ME
    [J]. ANALYTICAL CHEMISTRY, 1994, 66 (09) : 1369 - 1377
  • [7] Functional characterization of a conducting polymer-based immunoassay system
    Fare, TL
    Cabelli, MD
    Dallas, SM
    Herzog, DP
    [J]. BIOSENSORS & BIOELECTRONICS, 1998, 13 (3-4) : 459 - 470
  • [8] EVALUATION OF NAFION AS MEDIA FOR GLUCOSE-OXIDASE IMMOBILIZATION FOR THE DEVELOPMENT OF AN AMPEROMETRIC GLUCOSE BIOSENSOR
    FORTIER, G
    VAILLANCOURT, M
    BELANGER, D
    [J]. ELECTROANALYSIS, 1992, 4 (03) : 275 - 283
  • [9] Flow-through amperometric immunosensor: Fast 'sandwich' scheme immunoassay
    Ghindilis, AL
    Krishnan, R
    Atanasov, P
    Wilkins, E
    [J]. BIOSENSORS & BIOELECTRONICS, 1997, 12 (05) : 415 - 423
  • [10] Immunosensors: Electrochemical sensing and other engineering approaches
    Ghindilis, AL
    Atanasov, P
    Wilkins, M
    Wilkins, E
    [J]. BIOSENSORS & BIOELECTRONICS, 1998, 13 (01) : 113 - 131