VZL 2010, 79(3):105-110

Immobilization of Acetylcholinesterase and a Construction of Biochemical Biosensor for Identification of Toxic Organophosphates

Miroslav Pohanka ORCID...1,2, Oto Drobík3, Zuzana Křenková4, Jana ®ďárová Karasová ORCID...2, Jiří Cabal2, Kamil Kuča ORCID...1,2
1 Univerzita obrany, Fakulta vojenského zdravotnictví, Centrum pokročilých studií, Hradec Králové
2 Univerzita obrany, Fakulta vojenského zdravotnictví, katedra toxikologie, Hradec Králové
3 Univerzita Karlova v Praze, Farmaceutická fakulta v Hradci Králové, katedra farmaceutické chemie, Hradec Králové
4 Univerzita Pardubice, Fakulta chemicko-technologická, katedra biologických a biochemických věd, Pardubice

Biosenzory jsou analytická zařízení zaloľená na těsném spojení tzv. biorekogničního elementu a fyzikálně chemického převodníku. V této studii byl pouľit sítotiskový senzor s platinovou pracovní elektrodou jako převodník a acetylcholinesterasou jako biorekogničním elementem. Acetylcholinesterasa byla imobilizována rozdílným způsobem, včetně sol-gel technologie, imobilizace na grafitová nanovlákna, precipitace glutaraldehydem a zachycení do sférických grafitových mikročástic. Pro jednotlivé imobilizační postupy byla stanovena účinnost a diskutuje se o praktickém dopadu konstrukce biosenzoru.

Keywords: Biosenzor; Acetylcholinesterasa; Nervově paralytická látka; Detekce; Stanovení; Organofosfát

Biosensors are analytical devices based on tight junction of so-called biorecognition element and a physicochemical transducer. In this study, a screen printed sensor with platinum working electrode as the transducer and acetylcholinesterase as the biorecognition element were used. Acetylcholinesterase was immobilized in a different way including sol-gel technologies, immobilization on graphite nanofibers, precipitation with glutaraldehyde and interception into spherical graphite microparticles. Efficacy for given immobilization protocols was estimated, and practical impact of the study was discussed.

Keywords: Biosensor; Acetylcholinesterase; Nerve agents; Detection; Assay; Organophosphate

Received: January 11, 2010; Published: September 1, 2010  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Pohanka, M., Drobík, O., Křenková, Z., ®ďárová Karasová, J., Cabal, J., & Kuča, K. (2010). Immobilization of Acetylcholinesterase and a Construction of Biochemical Biosensor for Identification of Toxic Organophosphates. Vojenské Zdravotnické Listy, 79(3), 105-110
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. ALTERIIS, E., de - PARASCANDOLA, P. - PECORELLA, MA., et al. Effect of galatin - immobilization on the catalytic activity of enzymes and microbial cells. Biotechnol. Techniq., 1988, vol. 2, no. 3, p. 205-210. Go to original source...
    2. BUKOWSKA, B. - HUTNIK, K. 2,4-D and MCPA and their derivatives: effect on the activity of membrane erythrocytes acetylcholinesterase (in vitro). Pest. Biochem. Physiol., 2006, vol. 85, no. 3, p. 174-180. Go to original source...
    3. CHUVAILO, ON. - DZYADEVICH, SV. - ELSKAYA, AV., et al. Carbon fibre based microbiosensors for in vivo measurements of acetylcholine and choline. Biosens. Bioelectron., 2004, vol. 21, no. 1, p. 87-94. Go to original source... Go to PubMed...
    4. CIUCU, AA. - NEGULESCU, C. - BALDWIN, RP. Detection of pesticides using an amperometric biosensor based on ferophthalocyanine chemically modified carbon paste electrode and immobilized bienzymatic system. Biosens. Bioelectron., 2003, vol. 18, no. 2, p. 303-310. Go to original source... Go to PubMed...
    5. DOONG, RA. - TSAI, HC. Immobilization and characterization of sol-gel encapsulated acetylcholinesterase fiber-optic biosensor. Anal. Chim. Acta, 2001, vol. 434, no. 2, p. 239-246. Go to original source...
    6. DORČÁK, V. - ©ESTÁKOVÁ, I. Electrochemical behavior of phytochelatins and related peptides at the hanging mercury drop electrode in the presence of cobalt (II) ions. Bioelectrochemistry, 2006, vol. 68, no. 1, p. 14-21. Go to original source... Go to PubMed...
    7. FALASCA, C. - PERRIER, N. - MASSOULIE, J., et al. Determinants of the t peptide involved in folding, degradation, and secretion of acetylcholinesterase. J. Biol. Chem., 2005, vol. 280, no. 2, p. 878-886. Go to original source... Go to PubMed...
    8. FROST, RG. - MONTHONY, JF. - ENGELHORN, SC., et al. Covalent immobilization of proteins to N-hydroxysuccinimide ester derivatives of agarose: Effect of protein charge on immobilization. Biochim. Biophys. Acta, 1981, vol. 670, no. 2, p. 164-169. Go to original source... Go to PubMed...
    9. HARTNETT, AM. - INGERSOLL, CM. - BAKER, GA., et al. Kinetics and thermodynamics of free flavin and the flavin- based redox active site within glucose oxidase dissolved in solution or sequestered within a sol-gel derived glass. Anal. Chem., 1999, vol. 71, no. 1, p. 1215-1224. Go to original source... Go to PubMed...
    10. HENCH, LL. - WEST, JK. The sol-gel process. Chem. Rev., 1990, vol. 90, no. 1, p. 33-79. Go to original source...
    11. NOVAK, I. - SERUGA, M. - KOMORKSY-LOVRIC, S. Square-wave and cyclic voltammetry of epicatechin gallate on glassy carbon electrode. J. Electroanal. Chem., 2009, vol. 631, no. 1, p. 71-75. Go to original source...
    12. POHANKA, M. - ZBOŘIL, P. Amperometric biosensor for D-lactate assay. Food Technol. Biotechnol., 2008, vol. 46, no. 1, p. 107-110.
    13. POHANKA, M. - BANDOUCHOVA, H. - SOBOTKA, J., et al. Comparison of ferric reducing antioxidant power and square wave voltammetry for assay of low molecular weight antioxidants in blood plasma: Performance and comparison of methods. Sensors, 2009, vol. 9, no. 1, p. 9094-9103. Go to original source... Go to PubMed...
    14. RENAULT, L. - ESSONO, S. - JUIN, M., et al. Structural insights into AChE inhibition by monoclonal antibodies. Chem. Biol. Interact., 2005, vol. 157, no. 1, p. 397-400. Go to original source... Go to PubMed...
    15. SEKEROGLU, G. - FADILOGLU, S. - GOGUS, F. Immobilization and characterization of naringinase for the hydrolysis of naringin. Eur. Food Res. Technol., 2006, vol. 224, no.1, p. 55-60. Go to original source...
    16. SHAHROKHIAN, S. - BOZORGZADEH, S. Electrochemical oxidation of dopamine in the presence of sulfhydryl compounds: application to the square-wave voltammetric detection of penicillamine and cysteine. Electrochim. Acta, 2006, vol. 51, no. 20, p. 4271-4276. Go to original source...
    17. SOTIROPOULOU, S. - CHANIOTAKIS, NA. Tuning the sol-gel microenvironment for acetylcholinesterase encapsulation. Biomaterials, 2005, vol. 26, no. 33, p. 6771-6779. Go to original source... Go to PubMed...
    18. SUPALKOVA, V. - HUSKA, D. - DIOPAN, V., et al. Electroanalysis of plant thiols. Sensors, 2007, vol. 7, no. 1, p. 932-959. Go to original source...
    19. SINNER, EK. - RITZ, S. - NAUMANN, R., et al. Self-assembled tethered biomolecular lipid membranes. Adv. Clin. Chem., 2009, vol. 49, no. 1, p. 159-179. Go to original source... Go to PubMed...
    20. TANG, H. - CHEN, J. - YAO, S., et al. Amperometric glucose biosensor based on adsorption of glucose oxidase at platinum nanoparticle-modified carbon nanotube electrode. Anal. Biochem., 2004, vol. 331, no. 1, p. 89-97. Go to original source... Go to PubMed...
    21. TIMUR, S. - PAZARLIOGLU, N. - PILLOTON, R., et al. Detection of phenolic compounds by thick film sensors based on Pseudomonas putida. Talanta, 2003, vol. 61, no. 2, p. 87-93. Go to original source... Go to PubMed...
    22. YAZICI, MS. - SELMAN, JR. Dissolution of partially immersed nickel during in situ oxidation in molten carbonate: cyclic, stripping and square wave voltammetry measurements. J. Electroanal. Chem., 1998, vol. 457, no. 1, p. 89-97. Go to original source...

    Return to the content