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Progress in the development of immunoanalytical methods incorporating recombinant antibodies to small molecular weight biotoxins

机译:结合针对小分子生物毒素的重组抗体的免疫分析方法的开发进展

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Rapid immunoanalytical screening of food and environmental samples for small molecular weight (hapten) biotoxin contaminations requires the production of antibody reagents that possess the requisite sensitivity and specificity. To date animal-derived polyclonal (pAb) and monoclonal (mAb) antibodies have provided the binding element of the majority of these assays but recombinant antibodies (rAb) isolated from in vitro combinatorial phage display libraries are an exciting alternative due to (1) circumventing the need for experimental animals, (2) speed of production in commonly used in vitro expression systems and (3) subsequent molecular enhancement of binder performance. Short chain variable fragments (scFv) have been the most commonly employed rAb reagents for hapten biotoxin detection over the last two decades but antibody binding fragments (Fab) and single domain antibodies (sdAb) are increasing in popularity due to increased expression efficiency of functional binders and superior resistance to solvents. rAb-based immunochromatographic assays and surface plasmon resonance (SPR) biosensors have been reported to detect sub-regulatory levels of fungal (mycotoxins), marine (phycotoxins) and aquatic biotoxins in a wide range of food and environmental matrices, however this technology has yet to surpass the performances of the equivalent mAb- and pAb-based formats. As such the full potential of rAb technology in hapten biotoxin detection has yet to be achieved, but in time the inherent advantages of engineered rAb are set to provide the next generation of ultra-high performing binder reagents for the rapid and specific detection of hapten biotoxins.
机译:对食品和环境样品进行快速免疫分析筛查是否存在小分子(半抗原)生物毒素污染,需要生产具有所需灵敏度和特异性的抗体试剂。迄今为止,动物衍生的多克隆(pAb)和单克隆(mAb)抗体已提供了大多数这些测定的结合元件,但由于(1)规避,从体外组合噬菌体展示库中分离的重组抗体(rAb)是令人兴奋的选择。对实验动物的需求;(2)常用体外表达系统的生产速度;以及(3)结合剂性能的后续分子增强。在过去的二十年中,短链可变片段(scFv)已成为半抗原生物毒素检测中最常用的rAb试剂,但是由于功能结合剂的表达效率提高,抗体结合片段(Fab)和单域抗体(sdAb)的受欢迎程度日益提高以及优异的耐溶剂性。据报道,基于rAb的免疫色谱分析和表面等离振子共振(SPR)生物传感器可检测多种食品和环境基质中真菌(霉菌毒素),海洋(藻毒素)和水生生物毒素的亚调节水平,但是该技术尚未超越同等基于mAb和pAb的格式的性能。因此,rAb技术在半抗原生物毒素检测中的全部潜力尚未实现,但是随着时间的推移,经过工程改造的rAb的固有优势将被设置为提供下一代超高性能结合剂,用于快速,特异性地检测半抗原生物毒素。 。

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