Synthesis of Quenchbodies for One-Pot Detection of Stimulant Drug Methamphetamine

Abstract

The problem of illicit drug use and addiction is an escalating issue worldwide. As such, fast and precise detection methods are needed to help combat the problem. Herein, the synthesis method for an anti-methamphetamine Quenchbody (Q-body), a promising sensor for use in simple and convenient assays, has been described. The fluorescence intensity of the Q-body generated by two-site labeling of Escherichia coli produced anti-methamphetamine antigen-binding fragment (Fab) with TAMRA-C2-maleimide dyes increased 5.1-fold over background in the presence of a hydroxyl methamphetamine derivative, 3-[(2S)-2-(methylamino)propyl]phenol. This derivative has the closest structure to methamphetamine of the chemicals available for use in a laboratory. Our results indicate the potential use of this Q-body as a novel sensor for the on-site detection of methamphetamine, in such occasions as drug screening at workplace, suspicious substance identification, and monitoring patients during drug rehabilitation.

Nanobody Engineering: Computational Modelling and Design for Biomedical and Therapeutic Applications

“…Among these are the scFv-based Q-bodies that detect the antidepressant fluvoxamine [150] and the proinflammatory cytokine tumor necrosis factor alpha [153]. Additionally, there are Fab-based Q-bodies (also referred to as ultra Qbodies) that detect the highly-addictive psychostimulant methamphetamine [154], herbal cannabis [155] and amyloid β oligomers to aid the diagnosis of Alzheimer's disease [156].…”

Nanobody Engineering: Computational Modelling and Design for Biomedical and Therapeutic Applications

“…Among these are the scFv-based Q-bodies that detect the antidepressant fluvoxamine [148] and the proinflammatory cytokine tumor necrosis factor alpha [151]. Additionally, there are Fab-based Q-bodies (also referred to as ultra Qbodies) that detect the highly-addictive psychostimulant methamphetamine [152], herbal cannabis [153] and amyloid β oligomers to aid the diagnosis of Alzheimer's disease [154]. Finally, nanobody-based mini Q-bodies that detect the small hapten methotrexate [146], lysozyme [155] and recombinant human growth hormone and its isoforms [149] have been developed.…”

Generation of an scFv-type Quenchbody Against HER2 for Sensitive Detection of Human Breast Cancer

“…To date, different methods for generating Quenchbodies have been developed and separately used according to the research purposes and in an experimentally executable environment [12,17]. Among them, a maleimide-thiol reaction-based method has been used for post-labeling recombinant antibody fragments, expressed from E. coli [10,18,19]. Maleimide-thiol reaction is a click chemistry reaction, widely used for conjugating fluorescent dye to protein [20].…”