Microbead-assisted PDA sensor for the detection of genetically modified organisms

A simple and sensitive approach for the detection of marker protein, phosphinothricin acetyltransferase, from genetically modified crops was developed based on the colorimetric transition of polydiacetylene (PDA) vesicles in combination with silica microbeads. PDAs have attracted a great deal of interests as a transducing material due to their special features that allow colorimetric response to sensory signals, as well as their inherent simplicity. However, most PDA-based biosensors require additional analytical equipment such as a fluorescence microscope or UV–Vis spectrometer. In this study, we report a new approach to increase the degree of color transition by coupling antibody-conjugated PDA vesicles with silica microbeads in an effort to monitor the results with the unaided eye or simple RGB analysis. By immobilizing PDA vesicles on silica microbeads, we were able to overcome the disadvantages of colloidal PDA-based sensors and increase the degree of colorimetric changes in response to target molecules to a concentration as low as 20 nM. The additional stresses were given to PDA vesicles by antigen–antibody bridging of PDA vesicles coupled with microbeads, resulting in enhanced blue–red color transition. All the results showed that PDA vesicles in conjunction with silica microbeads will be a promising transducing material for the detection of target proteins in diagnostic and biosensing applications.

[1]  T. Schrader,et al.  Selective detection of catecholamines by synthetic receptors embedded in chromatic polydiacetylene vesicles. , 2005, Journal of the American Chemical Society.

[2]  Raymond C. Stevens,et al.  Coupling of an induced fit enzyme to polydiacetylene thin films: Colorimetric detection of glucose , 1997 .

[3]  Long Jiang,et al.  Chromatic immunoassay based on polydiacetylene vesicles. , 2004, Colloids and surfaces. B, Biointerfaces.

[4]  A. Roda,et al.  New trends in bioanalytical tools for the detection of genetically modified organisms: an update , 2008, Analytical and bioanalytical chemistry.

[5]  Long Jiang,et al.  Biosensor signal amplification of vesicles functionalized with glycolipid for colorimetric detection of Escherichia coli. , 2005, Journal of colloid and interface science.

[6]  H. Kuiper,et al.  Summary report of the ILSI Europe workshop on detection methods for novel foods derived from genetically modified organisms , 1999 .

[7]  A. Basu,et al.  Lipopolysaccharide identification with functionalized polydiacetylene liposome sensors. , 2004, Journal of the American Chemical Society.

[8]  Dong June Ahn,et al.  Effect of phospholipid insertion on arrayed polydiacetylene biosensors. , 2008, Colloids and surfaces. B, Biointerfaces.

[9]  A. Singh,et al.  One-step, chemisorbed immobilization of highly stable, polydiacetylenic phospholipid vesicles onto gold films. , 2001, Journal of the American Chemical Society.

[10]  Franco Pedreschi,et al.  A non-destructive digital imaging method to predict immobilized yeast-biomass , 2009 .

[11]  Laurence Meagher,et al.  Control over PEGylated-liposome aggregation by NeutrAvidin-biotin interactions investigated by photon correlation spectroscopy , 2002 .

[12]  B. A. Pindzola,et al.  Biosensing with polydiacetylene materials: structures, optical properties and applications. , 2007, Chemical communications.

[13]  R C Stevens,et al.  A 'litmus test' for molecular recognition using artificial membranes. , 1996, Chemistry & biology.

[14]  Sang Hoon Lee,et al.  Micropatterning of diacetylenic liposomes on glass surfaces , 2004 .

[15]  Dong June Ahn,et al.  Immobilized Polydiacetylene Vesicles on Solid Substrates for Use as Chemosensors , 2003 .

[16]  B. Tieke.  Polymerization of butadiene and butadiyne (diacetylene) derivatives in layer structures , 1985 .

[17]  B. A. Pindzola,et al.  Polydiacetylene Liposomes Attached to Glass Fibers for Fluorescent Bioassays , 2006 .

[18]  Yun Kyung Jung,et al.  Specific Colorimetric Detection of Proteins Using Bidentate Aptamer‐Conjugated Polydiacetylene (PDA) Liposomes , 2010 .

[19]  Deborah H. Charych,et al.  Polydiacetylene liposomes functionalized with sialic acid bind and colorimetrically detect influenza virus , 1995 .

[20]  Arno Schulz,et al.  Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants. , 2005, Regulatory toxicology and pharmacology : RTP.

[21]  T. Reuter,et al.  Investigations on Genetically Modified Maize (Bt-Maize) in Pig Nutrition: Chemical Composition and Nutritional Evaluation , 2002, Archiv fur Tierernahrung.

[22]  C. Carter,et al.  International Approval and Labeling Regulations of Genetically Modified Food in Major Trading Countries , 2006 .

[23]  Raz Jelinek,et al.  Biomolecular Sensing with Colorimetric Vesicles , 2007 .

[24]  M. Oh,et al.  Micro-patterned polydiacetylene vesicle chips for detecting protein-protein interactions , 2006 .

[25]  Raz Jelinek,et al.  Rapid Chromatic Detection of Bacteria by Use of a New Biomimetic Polymer Sensor , 2006, Applied and Environmental Microbiology.

[26]  Jong-Man Kim,et al.  A polydiacetylene-based fluorescent sensor chip. , 2005, Journal of the American Chemical Society.

[27]  David Zilberman,et al.  Regulating agricultural biotechnology : economics and policy , 2006 .

[28]  Zhanfang Ma,et al.  Colorimetric Detection of Escherichia coli by Polydiacetylene Vesicles Functionalized with Glycolipid , 1998 .

[29]  Gerhard Wegner,et al.  Topochemische Reaktionen von Monomeren mit konjugierten Dreifachbindungen / Tochemical Reactions of Monomers with conjugated triple Bonds , 1969 .