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Development of Protein A Functionalized Microcantilever Immunosensors for the Analyses of Small Molecules at Parts per Trillion Levels

机译:蛋白质A功能化的微悬臂免疫传感器的开发,用于在万亿分之一的水平上分析小分子。

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摘要

Development of microcantilever biosensors for small mol-necules was exemplified with the u0001-adrenergic agonist clen-nbuterol and the antibiotic chloramphenicol. In this paper,nantibody sulfhydrylation and protein A were used to modifynthe microcantilever Au surface, and the antibody activitiesnon the microcantilever were evaluated with direct competi-ntive enzyme-linked immunosorbent assay (dcELISA). Thenactivity of the antibodies immobilized on the microcanti-nlever via protein A was 1.7-fold of that via the sulfhydry-nlation reagent 2-iminothiolane hydrochloride. A micro-ncantilever immunosensor method with protein A as thenfunctionalization reagent was established to detect thenresidues of clenbuterol and chloramphenicol at limits ofndetection (LOD) of approximately 0.1 and 0.2 ng/mL,nrespectively. Such LODs were better than that of thencorresponding dcELISAs. The concentration of clen-nbuterol in a fortified feed sample detected with thenmicrocantilever immunosensor after thorough extractionnand purification agreed well with that detected with thendcELISA. Protein A showed to be simple and reproduciblenfor functionalization of the antibodies on the Au surfacenand, thus, has common application values in microcan-ntilever immunosensor development. The results suggestnthat microcantilever immunosensors be suitable for de-ntection of small molecules, and the assay sensitivity isnmainly related to the quality and activities of the antibodies.
机译:u0001-肾上腺素能激动剂clen-nbuterol和抗生素氯霉素证明了小分子微悬臂生物传感器的开发。本文利用抗体的巯基化和蛋白A修饰微悬臂梁的Au表面,并通过直接竞争酶联免疫吸附试验(dcELISA)评估了微悬臂梁的抗体活性。然后,通过蛋白质A固定在微抗体上的抗体的活性是通过巯基化试剂2-亚氨基硫杂环戊烷盐酸盐的抗体的活性的1.7倍。建立了以蛋白质A为功能化试剂的微悬臂免疫传感器方法,分别检测克伦特罗和氯霉素的残留量,其检出限(LOD)分别为约0.1和0.2 ng / mL。此类LOD优于相应的dcELISA。经过彻底提取和纯化后,再用微悬臂免疫传感器检测到的强化饲料样品中盐酸克伦特罗的浓度与使用dcELISA检测的浓度非常吻合。蛋白A对于在Au表面分子上的抗体功能化显示出简单且可重现的特点,因此在微悬臂免疫传感器的开发中具有共同的应用价值。结果表明,微悬臂梁式免疫传感器适用于小分子的检测,其检测灵敏度主要与抗体的质量和活性有关。

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  • 来源
    《Analytical Chemistry》 |2010年第2期|p.615-620|共6页
  • 作者单位

    College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, People’s Republic of China,and Key Laboratory of Mechanical Behavior and Design of Material of Chinese Academy of Sciences, University ofScience and Technology of China, Hefei 230027, People’s Republic of China;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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  • 入库时间 2022-08-17 13:36:36

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