J 2003

Development of piezoelectric immunosensors for competitive and direct determination of atrazine

PRIBYL, Jan, Maria HEPEL, Jan HALÁMEK and Petr SKLÁDAL

Basic information

Original name

Development of piezoelectric immunosensors for competitive and direct determination of atrazine

Authors

PRIBYL, Jan (203 Czech Republic), Maria HEPEL (840 United States of America), Jan HALÁMEK (203 Czech Republic) and Petr SKLÁDAL (203 Czech Republic, guarantor)

Edition

Sensors and Actuators B Chemical, Amsterdam (Holandsko), Elsevier, 2003, 0925-4005

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 2.391

RIV identification code

RIV/00216224:14310/03:00008696

Organization unit

Faculty of Science

UT WoS

000183039200054

Keywords in English

Quartz crystal microbalance; Immunosensor; Atrazine

Tags

Změněno: 14/5/2003 09:12, prof. RNDr. Petr Skládal, CSc.

Abstract

V originále

The improved highly sensitive piezoelectric immunosensor has been developed. The gold electrodes of the piezoelectric quartz crystals were modified with self-assembled layer of cysteamine and carboxyderivative of atrazine was covalently attached after activation with O-(N-succinimidyl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TSTU). The competitive immunoassay for atrazine was developed using anti-atrazine monoclonal antibody (MAb) D6F3. The limit of detection for atrazine was 0.025 ng/ml, the total time of analysis being 25 min. The repeated use of the immunosensors was possible after regeneration using pepsin at pH 2.0. The direct immunosensor for atrazine was constructed by oriented immobilization of anti-atrazine MAb to Protein A covalently attached to the gold surface activated with 3,3'-dithio-bis(propionic acid N-hydroxysuccinimide ester) (DTSP). The MAb-Protein A complex was stabilized by cross-linking with dimethyl pimelimidate. Thus developed immunosensor was able to specifically respond to atrazine present in solution, providing limit of detection of 1.5 ng/ml, one assay was completed within 10 min (5 min binding, 5 min washing of surface). The observed changes of frequency are expected to result from conformational changes of the immobilized antibody after formation of the immunocomplex with atrazine. Furthermore, kinetic studies were carried out in different arrangements, the obtained kinetic association equilibrium constant was 2.3x10^7 l mol-1 for interaction between immobilized antibody and atrazine in solution.

Links

OC 518.30, research and development project
Name: Sledování interakcí biomakromolekul s biovrstvami v různých konformačních stavech pomocí piezoelektrických biosensorů
Investor: Ministry of Education, Youth and Sports of the CR, Characterization of interactions of biomacromolecules with biolayers in different conformation states using piezoelectric biosensors