Polyelectrolyte Enzymatic Microdiagnosticum
The Production Technology and Properties of Polyelectrolyte Enzymatic Nano- and Microdiagnosticumes – a Novel Class of Biochemical Sensors
Tech Area / Field
- BIO-IND/Industrial Biotechnology/Biotechnology
3 Approved without Funding
Institute of Theoretical and Experimental Biophysics, Russia, Moscow reg., Puschino
- The Institute for Instrumentation in Biology of Russian Academy of Science, Russia, Moscow reg., Puschino
- Gent University / Faculteit Farmaceutische Wetenschappen, Belgium, Gent\nUniversity of Turku / Finnish-Russian Joint Biotechnology Laboratory, Finland, Turku\nJacobs University, Germany, Bremen\nLouisiana Tech University / Institute for Micromanufacturing, USA, LA, Ruston\nUniversity of Michigan / Materials Science and Biomedical Engineering, USA, MI, Ann Arbor
Project summaryThe aim of the Project.
Development of technology for fabrication of polyelectrolyte nano- and microcapsules containing enzymes. Determination of physical and chemical properties of the capsules. Elaboration of polyelectrolyte enzymatic microdiagnosticumes based on these capsules.
Polyelectrolyte nano- and microcapsules (PEN&MC) are made by layer-by-layer adsorption of oppositely charged polyelectrolytes onto disperse nano- and micro-sized particles followed by their degradation and removal. At present PEN&MC are under intensive use for fastly developing field of polymeric nanotechnology.
The main feature of these capsule’s shells is semipermeability: it is permeable for low molecular substances whils impermeab for high – molecular substances and aggregates. A number of properties result from this feature, what determines an importance of capsules for both fundamental and applied researches.
This project is aimed for fabrication of the polyelectrolyte microdiagnosticum based on single nano- or microsized capsule containing enzyme. Such a microreactor composed of multilayer semipermeable shell made of polyelectrolytes. Essence of this method is that while placed into the liquid under the study this microreactor is capable to detect concentration of investigating substance by means of quantity of product of enzymatic reaction or by change of enzyme’s activity. The analyzable substance could be substrate or activator or inhibitor of enzyme use microdiagnosticum.
Preliminary results have showed that our methods to determine the concentrations of analyzable substances in the biological liquids (blood, urine, saliva, milk, and others) developed by our team have the following advantages over existing clinical - biochemical enzymatic methods:
- The encapsulated enzyme retains its activity during several months, whereas activity of the enzyme in the solution is decreased nearly to zero in several days.
- The encapsulated enzyme fully retains its activity in the biological fluids containing proteinases, which eliminates the necessity for their removal from investigating medium under analysis. Microdiagnosticum permits to make analyzes in-situ in biological liquid without its preliminary fractionation.
- This method exhibits possibility of multiple usage of the microdiagnosticum, which provides a minimization of enzyme’s quantity. It is necessary to admit that single polyelectrolyte capsule contain several picogramms of enzyme.
- The proposed method allows to determine concentration of the analyzable substance using only few capsules or even single capsule in the case when capsule’s shell labeling for visualization. Labelling can be done by fluorescent or radioactive probe or when interaction of enzyme with analyzable substance results in intensive chemiluminescence.
The executors of the project have a wide range of scientific knowledge and a competent experience in the polyelectrolyte nanotechnology. The research team includes 7 doctors of science, 4 Ph.Ds, post-graduates and students.
Since 2002 the research on PE&MC became a major direction for executors of the project. The main task was to develop practical usage of PE&MC in nanotechnology and medicine. The results of theoretical and applied research have been published in 19 articles in scientific journals. The executors’ list of publications on the subject of the project is attached. Materials and technical basis for fabrication and elaboration PE&MC is fully available and will provide reliable background to the successful performance of project.