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Detection of E. coli labeled with metal-conjugated antibodies using lateral-flow assay and laser-induced breakdown spectroscopy.
Gondhalekar, Carmen; Biela, Eva; Rajwa, Bartek; Bae, Euiwon; Patsekin, Valery; Sturgis, Jennifer; Reynolds, Cole; Doh, Iyll-Joon; Diwakar, Prasoon; Stanker, Larry; Zorba, Vassilia; Mao, Xianglei; Russo, Richard; Robinson, J Paul.
Afiliação
  • Gondhalekar C; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, 47907, USA.
  • Biela E; Department of Basic Medical Sciences, Purdue University, West Lafayette, IN, 47907, USA.
  • Rajwa B; Bindley Bioscience Center, Discovery Park, Purdue University, West Lafayette, IN, 47907, USA.
  • Bae E; School of Mechanical Engineering, Applied Optics Laboratory, Purdue University, West Lafayette, IN, 47907, USA.
  • Patsekin V; Department of Basic Medical Sciences, Purdue University, West Lafayette, IN, 47907, USA.
  • Sturgis J; Department of Basic Medical Sciences, Purdue University, West Lafayette, IN, 47907, USA.
  • Reynolds C; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, 47907, USA.
  • Doh IJ; School of Mechanical Engineering, Applied Optics Laboratory, Purdue University, West Lafayette, IN, 47907, USA.
  • Diwakar P; Mechanical Engineering Department, South Dakota School of Mines & Technology, Rapid City, SD, 57701, USA.
  • Stanker L; United States Department of Agriculture, USDA Research Services, Albany, CA, 94706, USA.
  • Zorba V; Energy Technologies Area, Lawrence Berkeley National Laboratories, Berkeley, CA, 94720, USA.
  • Mao X; Energy Technologies Area, Lawrence Berkeley National Laboratories, Berkeley, CA, 94720, USA.
  • Russo R; Energy Technologies Area, Lawrence Berkeley National Laboratories, Berkeley, CA, 94720, USA.
  • Robinson JP; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, 47907, USA. jpr@flowcyt.cyto.purdue.edu.
Anal Bioanal Chem ; 412(6): 1291-1301, 2020 Feb.
Article em En | MEDLINE | ID: mdl-31989196
This study explores the adoption of laser-induced breakdown spectroscopy (LIBS) for the analysis of lateral-flow immunoassays (LFIAs). Gold (Au) nanoparticles are standard biomolecular labels among LFIAs, typically detected via colorimetric means. A wide diversity of lanthanide-complexed polymers (LCPs) are also used as immunoassay labels but are inapt for LFIAs due to lab-bound detection instrumentation. This is the first study to show the capability of LIBS to transition LCPs into the realm of LFIAs, and one of the few to apply LIBS to biomolecular label detection in complete immunoassays. Initially, an in-house LIBS system was optimized to detect an Au standard through a process of line selection across acquisition delay times, followed by determining limit of detection (LOD). The optimized LIBS system was applied to Au-labeled Escherichia coli detection on a commercial LFIA; comparison with colorimetric detection yielded similar LODs (1.03E4 and 8.890E3 CFU/mL respectively). Optimization was repeated with lanthanide standards to determine if they were viable alternatives to Au labels. It was found that europium (Eu) and ytterbium (Yb) may be more favorable biomolecular labels than Au. To test whether Eu-complexed polymers conjugated to antibodies could be used as labels in LFIAs, the conjugates were successfully applied to E. coli detection in a modified commercial LFIA. The results suggest interesting opportunities for creating highly multiplexed LFIAs. Multiplexed, sensitive, portable, and rapid LIBS detection of biomolecules concentrated and labeled on LFIAs is highly relevant for applications like food safety, where in-field food contaminant detection is critical. Graphical abstract.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Análise Espectral / Escherichia coli / Lasers / Metais / Anticorpos Antibacterianos Tipo de estudo: Diagnostic_studies Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Análise Espectral / Escherichia coli / Lasers / Metais / Anticorpos Antibacterianos Tipo de estudo: Diagnostic_studies Idioma: En Ano de publicação: 2020 Tipo de documento: Article