Halámek Jan, Makower Alexander, Knösche Kristina, Skládal Petr, Scheller Frieder W
Department of Analytical Biochemistry, Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Golm, Germany.
Talanta. 2005 Jan 30;65(2):337-42. doi: 10.1016/j.talanta.2004.07.008.
We report here the development of piezoelectric affinity sensors for cocaine and cholinesterase inhibitors based on the formation of affinity complexes between an immobilized cocaine derivative and an anti-cocaine antibody or cholinesterase. For both binding reactions benzoylecgonine-1,8-diamino-3,4-dioxaoctane (BZE-DADOO) was immobilized on the surface of the sensor. For immobilization, pre-conjugated BZE-DADOO with 11-mercaptomonoundecanoic acid (MUA) via 2-(5-norbornen-2,3-dicarboximide)-1,1,3,3-tetramethyluronium-tetrafluoroborate (TNTU) allowed the formation of a chemisorbed monolayer on the piezosensor surface. The detection of cocaine was based on a competitive assay. The change of frequency measured after 300s of the binding reaction was used as the signal. The maximum binding of the antibody resulted in a frequency decrease of 35Hz (with an imprecision 3%, n = 3) while the presence of 100pmoll(-1) cocaine decreased the binding by 11%. The limit of detection was consequently below 100pmoll(-1) for cocaine. The total time of one analysis was 15min. This BZE-DADOO-modified sensor was adapted for the detection of organophosphates. BZE-DADOO - a competitive inhibitor - served as binding element for cholinesterase in a competitive assay.
我们在此报告基于固定化可卡因衍生物与抗可卡因抗体或胆碱酯酶之间形成亲和复合物而开发的用于检测可卡因和胆碱酯酶抑制剂的压电亲和传感器。对于这两种结合反应,均将苯甲酰芽子碱 - 1,8 - 二氨基 - 3,4 - 二氧杂辛烷(BZE - DADOO)固定在传感器表面。为了进行固定,通过2 - (5 - 降冰片烯 - 2,3 - 二甲酰亚胺) - 1,1,3,3 - 四甲基脲四氟硼酸盐(TNTU)使预共轭的BZE - DADOO与11 - 巯基十一烷酸(MUA)形成化学吸附单层,从而固定在压电传感器表面。可卡因的检测基于竞争性分析。将结合反应300秒后测得的频率变化用作信号。抗体的最大结合导致频率下降35Hz(不精密度为3%,n = 3),而存在100pmol l⁻¹可卡因时,结合减少了11%。因此,可卡因的检测限低于100pmol l⁻¹。一次分析的总时间为15分钟。这种BZE - DADOO修饰的传感器适用于有机磷的检测。BZE - DADOO(一种竞争性抑制剂)在竞争性分析中用作胆碱酯酶的结合元件。