Immobkilisation of engineered molecules on electrodes and optical surfaces

J. Maly, E. Illiano, M. Sabato, M. De Francesco, V. Pinto, A. Masci, D. Masci, J. Masojidek, M. Sugiura, R. Franconi, R. Pilloton

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Abstract

Monolayers of genetically modified proteins with an hexahistidine tag, (His)6, were obtained by using a Ni-NTA chelator synthesized on gold-sputtered surfaces (via sulphide bonds), or on gold and graphite (via sililating agents) working electrodes of screen-printed devices.Two kinds of proteins were produced and purified for this study: (a) a recombinant antibody, derived from the 'single-chain Fv' (scFv) format, and (b) a photosystem II (PSII) core complex isolated from the mutant strain CP43-H of the thermophilic cyanobacterium Synechococcus elongatus. An scFv previously isolated from a synthetic 'phage display' library was further engineered with an alkaline phosphatase activity genetically added between the carboxy-terminal of the scFvs and the (His)6to allow direct measurement of immobilisation. Renewable specific binding of (His)6proteins to gold and graphite surfaces and fast and sensitive electrochemical or optical detection of analytes were obtained. Additionally, "on chip" protein preconcentration was conveniently achieved for biosensing purposes, starting from crude unpurified extracts and avoiding protein purification steps. © 2002 Elsevier Science B.V. All rights reserved.
Original languageEnglish
Pages (from-to)257 - 261
Number of pages5
JournalMaterials Science and Engineering C
Volume22
Issue number2
DOIs
Publication statusPublished - 1 Dec 2002

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All Science Journal Classification (ASJC) codes

  • Materials Science(all)
  • Condensed Matter Physics
  • Mechanics of Materials
  • Mechanical Engineering

Cite this

Maly, J., Illiano, E., Sabato, M., De Francesco, M., Pinto, V., Masci, A., ... Pilloton, R. (2002). Immobkilisation of engineered molecules on electrodes and optical surfaces. Materials Science and Engineering C, 22(2), 257 - 261. https://doi.org/10.1016/S0928-4931(02)00177-7