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

Analytical and Bioanalytical Chemistry volume 400, pages 777–785 (2011)Cite this article

Abstract

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.

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Abbreviations

DMPC:

1,2-Dimyristoyl phosphatidyl choline

EDC:

1-Ethyl-3-(3-dimethyl amino-propyl)carbodiimide

GMO:

Genetically modified organisms

NHS:

N-hydroxysuccinimide

PAT:

Phosphinothricin acetyltransferase

PDA:

Polydiacetylene

TCDA:

10,12-Tricosadiynoic acid

References

  1. Wegner G (1969) Topochemical reactions of monomers with conjugated triple bonds I. Polymerization of derivatives of 2,4-hexadiyne-1,6-diols in the crystalline state. A Naturforschung Teil B 24(7):824–832

    CAS  Google Scholar 

  2. Tieke B (1985) Polymerization of butadiene and butadiyne (diacetylene) derivatives in layer structures. Adv Polym Sci 71:79–151

    CAS  Google Scholar 

  3. Jelinek R, Kolusheva S (2007) Biomolecular sensing with colorimetric vesicles. Top Curr Chem 277:155–180

    Article  CAS  Google Scholar 

  4. Y-l Su, J-r L, Jiang L, Cao J (2005) Biosensor signal amplification of vesicles functionalized with glycolipid for colorimetric detection of Escherichia coli. J Colloid Interface Sci 284(1):114–119

    Article  Google Scholar 

  5. Silbert L, Ben Shlush I, Israel E, Porgador A, Kolusheva S, Jelinek R (2006) Rapid chromatic detection of bacteria by use of a new biomimetic polymer sensor. Appl Environ Microbiol 72(11):7339–7344

    Article  CAS  Google Scholar 

  6. Ma Z, Li J, Liu M, Cao J, Zou Z, Tu J, Jiang L (1998) Colorimetric detection of Escherichia coli by polydiacetylene vesicles functionalized with glycolipid. J Am Chem Soc 120(48):12678–12679

    Article  CAS  Google Scholar 

  7. Reichert A, Nagy JO, Spevak W, Charych D (1995) Polydiacetylene liposomes functionalized with sialic acid bind and colorimetrically detect influenza virus. J Am Chem Soc 117(2):829–830

    Article  CAS  Google Scholar 

  8. Charych D, Cheng Q, Reichert A, Kuziemko G, Stroh M, Nagy JO, Spevak W, Stevens RC (1996) A ‘litmus test’ for molecular recognition using artificial membranes. Chem Biol 3(2):113–120

    Article  CAS  Google Scholar 

  9. Kolusheva S, Molt O, Herm M, Schrader T, Jelinek R (2005) Selective detection of catecholamines by synthetic receptors embedded in chromatic polydiacetylene vesicles. J Am Chem Soc 127(28):10000–10001

    Article  CAS  Google Scholar 

  10. Cheng Q, Stevens RC (1997) Coupling of an induced fit enzyme to polydiacetylene thin films: colorimetric detection of glucose. Adv Mater 9(6):481–483

    Article  CAS  Google Scholar 

  11. Rangin M, Basu A (2004) Lipopolysaccharide identification with functionalized polydiacetylene liposome sensors. J Am Chem Soc 126(16):5038–5039

    Article  CAS  Google Scholar 

  12. Stanish I, Santos JP, Singh A (2001) One-step, chemisorbed immobilization of highly stable, polydiacetylenic hospholipid vesicles onto gold films. J Am Chem Soc 123(5):1008–1009

    Article  CAS  Google Scholar 

  13. Kim J-M, Ji E-K, Woo SM, Lee H, Ahn DJ (2003) Immobilized polydiacetylene vesicles on solid substrates for use as chemosensors. Adv Mater 15(13):1118–1121

    Article  CAS  Google Scholar 

  14. Shim HY, Lee SH, Ahn DJ, Ahn KD, Kim JM (2004) Micropatterning of diacetylenic liposomes on glass surfaces. Mater Sci Eng C 24(1–2):157–161

    Article  Google Scholar 

  15. Kim JM, Lee YB, Yang DH, Lee JS, Lee GS, Ahn DJ (2005) A polydiacetylene-based fluorescent sensor chip. J Am Chem Soc 127(50):17580–17581

    Article  CAS  Google Scholar 

  16. Reppy MA, Pindzola BA (2006) Polydiacetylene liposomes attached to glass fibers for fluorescent bioassays. Mater Res Soc Symp Proc 942:W13_10

    Article  Google Scholar 

  17. Just RE, Alston JM, Zilberman D, Carter C, Gruère G (2006) International approval and labeling regulations of genetically modified food in major trading countries. In: Zilberman D (ed) Regulating agricultural biotechnology: economics and policy, vol 30, Natural resource management and policy. Springer, New York, pp 459–480

    Chapter  Google Scholar 

  18. Michelini E, Simoni P, Cevenini L, Mezzanotte L, Roda A (2008) New trends in bioanalytical tools for the detection of genetically modified organisms: an update. Anal Bioanal Chem 392(3):355–367

    Article  CAS  Google Scholar 

  19. Herouet C, Esdaile DJ, Mallyon BA, Debruyne E, Schulz A, Currier T, Hendrickx K, van der Klis R-J, Rouan D (2005) Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants. Regul Toxicol Pharmacol 41(2):134–149

    Article  CAS  Google Scholar 

  20. Reuter T, Aulrich K, Berk A, Flachowsky G (2002) Investigations on genetically modified maize (Bt-maize) in pig nutrition: chemical composition and nutritional evaluation. Arch Anim Nutr 56(1):23–31

    Article  CAS  Google Scholar 

  21. Y-l Su, J-r L, Jiang L (2004) Chromatic immunoassay based on polydiacetylene vesicles. Colloids Surf, B 38(1–2):29–33

    Google Scholar 

  22. Kim K-W, Choi H, Lee GS, Ahn DJ, Oh M-K (2006) Micro-patterned polydiacetylene vesicle chips for detecting protein-protein interactions. Macromol Res 14:483–485

    CAS  Google Scholar 

  23. Reppy MA, Pindzola BA (2007) Biosensing with polydiacetylene materials: structures, optical properties and applications. Chem Commun (42):4317–4338

  24. Kim K-W, Choi H, Lee GS, Ahn DJ, Oh M-K (2008) Effect of phospholipid insertion on arrayed polydiacetylene biosensors. Colloids Surf, B 66(2):213–217

    Article  CAS  Google Scholar 

  25. Acevedo CA, Skurtys O, Young ME, Enrione J, Pedreschi F, Osorio F (2009) A non-destructive digital imaging method to predict immobilized yeast-biomass. LWT Food Sci Technol 42(8):1444–1449

    Article  CAS  Google Scholar 

  26. Vermette P, Taylor S, Dunstan D, Meagher L (2001) Control over PEGylated-liposome aggregation by NeutrAvidin-Biotin interactions investigated by photon correlation spectroscopy. Langmuir 18(2):505–511

    Article  Google Scholar 

  27. Jung YK, Kim TW, Park HG, Soh HT (2010) Specific colorimetric detection of proteins using bidentate aptamer-conjugated polydiacetylene (PDA) liposomes. Adv Funct Mater 20(18):3092–3097

    Article  CAS  Google Scholar 

  28. Shim Y-Y, Shin W-S, Moon G-S, Kim K-H (2007) Quantitative analysis of phosphinothricin-n-acetyltransferase in genetically modified herbicide tolerant pepper by an enzyme-linked immunosorbent assay. J Microbiol Biotechnol 17(4):681–684

    CAS  Google Scholar 

  29. Yates K (ed) (1999) Detection methods for novel foods derived from genetically modified organisms. ILSI Europe, Brussels

    Google Scholar 

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Acknowledgement

This work was supported by a grant (Grant No. 20080401034032) from BioGreen 21 program, Rural Development Administration, Republic of Korea.

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Affiliations

  1. Institute of Life Sciences and Resources & Department of Food Science and Biotechnology, College of Life Sciences, Kyung Hee University, Yongin, 446-701, South Korea

    Min-Cheol Lim, Yeo-Jae Shin, Hae-Yeong Kim & Young-Rok Kim

  2. Department of Biological Engineering, Inha University, Incheon, 402-751, South Korea

    Tae-Joon Jeon

Authors
  1. Min-Cheol Lim
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  2. Yeo-Jae Shin
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  3. Tae-Joon Jeon
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  4. Hae-Yeong Kim
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  5. Young-Rok Kim
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Correspondence to Young-Rok Kim.

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Lim, MC., Shin, YJ., Jeon, TJ. et al. Microbead-assisted PDA sensor for the detection of genetically modified organisms. Anal Bioanal Chem 400, 777–785 (2011). https://doi.org/10.1007/s00216-011-4832-7

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  • DOI: https://doi.org/10.1007/s00216-011-4832-7

Keywords

  • Colorimetric sensor
  • Polydiacetylene
  • Silica microbead
  • PAT
  • GMO