Physical and chemical characterization of enolase immobilized polydiacetylene Langmuir–Blodgett film
| Autor: | Gotam K. Jarori, Shilpa N. Sawant, K. Sadagopan, S.K. Kulshreshtha |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Chemistry
Enolase Metals and Alloys Substrate (chemistry) Condensed Matter Physics HEXA Langmuir–Blodgett film Surfaces Coatings and Films Electronic Optical and Magnetic Materials Covalent bond Materials Chemistry Organic chemistry Electrical and Electronic Engineering Instrumentation Biosensor Magnesium ion Histidine Nuclear chemistry |
| Zdroj: | Sensors and Actuators B: Chemical. 115:526-533 |
| ISSN: | 0925-4005 |
| DOI: | 10.1016/j.snb.2005.10.031 |
| Popis: | Hexa histidine tagged recombinant Plasmodium falciparum enolase (His6-Pfen) was covalently immobilized on a Langmuir–Blodgett film of a self assembled mixture of 10,12-pentacosadiynoic acid and its N-succinimidyl ester derivative (PDA LB-film). The film was polymerized with a UV-lamp at 254 nm to obtain a blue coloured, protein hooked polydiacetylene film. Atomic force microscopy (AFM) was used to characterize the surface morphology of the protein-immobilized film. The colorimetric response (CR) of the His6-Pfen hooked PDA LB-film to 2-phosphoglyceric acid (2-PGA), the substrate of enolase, in the presence of magnesium ions was studied spectrophotometrically. The CR of glutathione S-transferasePfen (GST-Pfen) immobilized film prepared by using similar procedure was also examined. The results suggest that binding of Mg (II) to the enzyme facilitates the interaction of the enzyme with 2-phosphoglycerate. This ligand-binding event could be detected by an observed increase in colorimetric response of the film by ∼10%. Thus the incorporation of enolase on a PDA film resulted in the formation of a novel material, which can serve as a biosensor. © 2005 Elsevier B.V. All rights reserved. |
| Databáze: | OpenAIRE |
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