| Titolo | Immobkilisation of engineered molecules on electrodes and optical surfaces |
|---|---|
| Tipo di pubblicazione | Articolo su Rivista peer-reviewed |
| Anno di Pubblicazione | 2002 |
| Autori | Maly, J., Illiano E., Sabato M., De Francesco M., Pinto V., Masci A., Masci D., Masojidek J., Sugiura M., Franconi Rosella, and Pilloton R. |
| Rivista | Materials Science and Engineering C |
| Volume | 22 |
| Paginazione | 257-261 |
| ISSN | 09284931 |
| Parole chiave | alkaline 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. |
| Note | cited By 21 |
| URL | https://www.scopus.com/inward/record.uri?eid=2-s2.0-0036920926&doi=10.1016%2fS0928-4931%2802%2900177-7&partnerID=40&md5=ec141156ed2a8d43b4bd3e70211f5ddb |
| DOI | 10.1016/S0928-4931(02)00177-7 |
| Citation Key | Maly2002257 |
