Immobkilisation of engineered molecules on electrodes and optical surfaces

TitleImmobkilisation of engineered molecules on electrodes and optical surfaces
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2002
AuthorsMaly, J., Illiano E., Sabato M., De Francesco M., Pinto V., Masci A., Masci D., Masojidek J., Sugiura M., Franconi Rosella, and Pilloton R.
JournalMaterials Science and Engineering C
Volume22
Pagination257-261
ISSN09284931
Keywordsalkaline phosphatase, bacterium mutant, chelating agent, Chelation, Electrodes, enzyme activity, Gold, Graphite, histidine, Monolayers, Mutagens, Nickel, nitrilotriacetic acid, Optical surface, Photosystem II, Protein, protein engineering, protein purification, Proteins, single chain fragment variable antibody, surface property, thermophilic bacterium
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)6 to allow direct measurement of immobilisation. Renewable specific binding of (His)6 proteins 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.

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URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-0036920926&doi=10.1016%2fS0928-4931%2802%2900177-7&partnerID=40&md5=ec141156ed2a8d43b4bd3e70211f5ddb
DOI10.1016/S0928-4931(02)00177-7