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Mirsky, Vladimir M. ; Mass, Markus ; Krause, Christian ; Wolfbeis, Otto S.

Capacitive Approach To Determine Phospholipase A2 Activity toward Artificial and Natural Substrates

Mirsky, Vladimir M., Mass, Markus, Krause, Christian und Wolfbeis, Otto S. (1998) Capacitive Approach To Determine Phospholipase A2 Activity toward Artificial and Natural Substrates. Analytical Chemistry 70 (17), S. 3674-3678.

Veröffentlichungsdatum dieses Volltextes: 18 Apr 2011 13:14
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.20456


Zusammenfassung

A capacitive approach has been employed to develop a novel method to determine phospholipase activity. The sensing electrodes have a structure like Au/S(CH2)17CH3/substrate/electrolyte. Hydrolysis of the substrate, mediated by phospholipase A2, leads to the formation of water-soluble products from the insoluble substrate. This results in desorption of these products into aqueous phase and ...

A capacitive approach has been employed to develop a novel method to determine phospholipase activity. The sensing electrodes have a structure like Au/S(CH2)17CH3/substrate/electrolyte. Hydrolysis of the substrate, mediated by phospholipase A2, leads to the formation of water-soluble products from the insoluble substrate. This results in desorption of these products into aqueous phase and corresponding increase of the electrode capacitance. The requirement of high water solubility of the reaction products can be achieved in two ways. In the first, short-chain phospholipids are used as the substrate, in which case, water-soluble products are formed and no additional reagents are required to promote desorption of these products. The sensors prepared by this strategy provide sensitive qualitative detection of phospholipases. The second way is based on the use of a water-soluble acceptor (for example, β-cyclodextrin) to solubilize the products of hydrolysis. It allows semiquantitative detection of phospholipase activity toward long-chain natural substrates. The reaction kinetics for this case was found to be monoexponential and linearly dependent on the phospholipase concentration. The detection limit of this method, as tested with phospholipase A2 from bee venom and soy bean lecithin as the substrate, is 0.5 ng/mL (500 μunits/mL).



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Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftAnalytical Chemistry
Verlag:American Chemical Society
Band:70
Nummer des Zeitschriftenheftes oder des Kapitels:17
Seitenbereich:S. 3674-3678
Datum1998
InstitutionenChemie und Pharmazie > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Antje J. Bäumner, ehemals Prof. Wolfbeis)
Identifikationsnummer
WertTyp
10.1021/ac980102wDOI
Dewey-Dezimal-Klassifikation500 Naturwissenschaften und Mathematik > 540 Chemie
StatusVeröffentlicht
BegutachtetUnbekannt / Keine Angabe
An der Universität Regensburg entstandenUnbekannt / Keine Angabe
Dokumenten-ID20456

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