TITLE:
Enzyme Substrates Protective Encapsulation within Polymeric Microspheres
AUTHORS:
Joanna Zajda, Agnieszka M. Jastrzębska, Andrzej R. Olszyna, Elżbieta Malinowska, Agata Michalska
KEYWORDS:
Enzyme Substrate Encapsulation; BSA; Lipase; Polyacrylate Microspheres
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.4 No.9,
September
4,
2013
ABSTRACT: Determination of enzymes
activity is an important task of analytical and biomedical oriented
fluorimetry. Despite of a long track
record of application, there is still some room for improvements in this field.
In the case of hydrolases, spontaneous decomposition of substrate leads to substantial errors in
determination of enzyme activity. An innovative and effective approach is
proposed allowing protection of enzyme substrate within the lipophilic moiety
of polyacrylate microspheres. It is shown that the introduction of substrate into
microspheres is not only an effective method of prevention of unwanted spontaneous process, but also
does not disturb the availability of substrate for enzymatic hydrolysis. The
effect of presence of proteins in the sample on fluorimetric responses was
studied. In contrary to previous studies related to application of lipophilic
polymers in biomedical analysis, it is shown that the presence of bovine serum
albumins enhances the sensitivity of fluorimetric determination. It is shown
that this surprising effect is related to adsorption of proteins on the
microspheres surface and change of surface charge of polymer.