Characterization of immobilization methods for African swine fever virus protein and antibodies with a piezoelectric immunosensor

被引:45
作者
Uttenthaler, E [1 ]
Kosslinger, C [1 ]
Drost, S [1 ]
机构
[1] Fraunhofer Inst Solid State Technol, D-80686 Munich, Germany
关键词
African swine fever; immobilization; immunosensor; quartz crystal microbalance;
D O I
10.1016/S0956-5663(98)00089-X
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A direct piezoelectric flow injection analysis immunoassay for the detection of African Swine Fever virus and antibodies is presented. The peptide-specific monoclonal antibody 18BG3 and the virus protein 73 were used for detection with a quartz crystal microbalance. Accumulation of the analyte on the surface of this mass-sensitive biosensor resulted in a shift of the resonant frequency. Highly selective receptor layers were applied on the sensing electrode of the quartz crystal for detection of the complementary analyte. Different immobilization methods proved to be appropriate for coating of the monoclonal antibody 18BC3. A quartz crystal covalently coated with the antibody 18BG3 detected virus protein VP73 samples more than 20 times and was stable for more than 30 days. The coating of virus protein was performed by physisorption. A sensor with a virus protein receptor layer detected antibody 18BG3 samples 10 times within one day. The sensor device was able to perform one measurement cycle including blocking and regeneration within 30 min. With the help of a suitable carrier liquid, measurements with serum samples were performed. The calibration curves for measurements in buffer and in serum could be determined and the detection limits for virus protein detection were 0.31 and 1 mu g/ml, and for antibody detection 0.1 and 0.2 mu g/ml respectively. (C) 1998 Elsevier Science S.A. All rights reserved.
引用
收藏
页码:1279 / 1286
页数:8
相关论文
共 20 条
[1]   HIV SEROLOGY USING PIEZOELECTRIC IMMUNOSENSORS [J].
ABERL, F ;
WOLF, H ;
KOSSLINGER, C ;
DROST, S ;
WOIAS, P ;
KOCH, S .
SENSORS AND ACTUATORS B-CHEMICAL, 1994, 18 (1-3) :271-275
[2]  
AHLUWALIA A, 1991, BIOSENS BIOELECTRON, V7, P207
[3]  
HARTMEIER W, 1986, IMMOBILISIERTE BIOKA
[4]  
Hermanson G.T., 1992, Immobilized affinity ligand techniques
[5]  
HESS WR, 1981, ADV VET SCI COMP MED, V25, P39
[6]   LABELING OF COLLOIDAL GOLD WITH PROTEIN - A QUANTITATIVE STUDY USING BETA-LACTOGLOBULIN [J].
HORISBERGER, M ;
VAUTHEY, M .
HISTOCHEMISTRY, 1984, 80 (01) :13-18
[7]  
KANAZAWA KK, 1985, ANAL CHIM ACTA, V175, P99
[8]  
KOSSLINGER C, 1995, SENSOR ACTUAT B-CHEM, V24, P107, DOI 10.1016/0925-4005(95)85023-6
[9]   QUARTZ-CRYSTAL MICROBALANCE FOR IMMUNOSENSING [J].
KOSSLINGER, C ;
DROST, S ;
ABERL, F ;
WOLF, H .
FRESENIUS JOURNAL OF ANALYTICAL CHEMISTRY, 1994, 349 (05) :349-354
[10]   3-DIMENSIONAL STRUCTURES OF AVIDIN AND THE AVIDIN-BIOTIN COMPLEX [J].
LIVNAH, O ;
BAYER, EA ;
WILCHEK, M ;
SUSSMAN, JL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (11) :5076-5080