Development of biosensors for analytes of high social interest based on advanced nano/biotechonological approaches.

Project leaders

Sabato Dauria, Konstantin Turoverov

Agreement

RUSSIA - RAS old - Russian Academy of Sciences old

Call

CNR/RAS 2011-2013

Department

Life Sciences

Thematic area

Biomedical sciences

Status of the project

Extended

Report for renewal

c-documents-and-settings-tino-desktop-report-cnr-ras.pdf

Research proposal

There is a substantial word-wide effort to develop non-invasive and minimally invasive methods for frequent and/or continuous monitoring of clinical analytes in blood or interstitial fluids. In addition, several research groups are also working on the development of biosensors for sensing analytes of high environmental interest, such as heavy metals, antibiotics, etc. A wide variety of methods potentially useful to measure the binding of analytes have been tested, including optical rotation, near-infrared absorbance, Raman scattering, as well as the design and synthesis of many chemical fluorescence probes. Fluorescence detection is the dominant analytical tool in medical testing, biotechnology and drug discovery. Starting in 1980s fluorescence probes for specific analytes become available, and some of these sensing fluorophores are relatively simple. However, the molecular complexity of the probes quickly increases if one requires analyte binding to cause a spectral change. For example, the fluorophores specific for calcium are structurally complex and only a few display spectral shifts upon binding calcium. As consequence, the development of specific sensors for biochemically relevant analytes is even more challenging. Even if a suitable structure could be designed and synthesized, there is no guarantee that the final molecule will display a spectral change, adequate water solubility, and a suitable affinity constant. To circumvent these difficulties, the use of proteins and enzymes as specific sensors for biochemical analytes has become a priority of the modern biotechnology. In this project we propose to use the D-serine dehydratase as specific probe for the design and realization of an advanced optical biosensor for the detection of D-serine in serum and/or interstitial fluids. A D-serine dehydratase has been already isolated from S. cerevisiae by and available in Dr. D'Auria's lab. The goal of our project is to over-produce this enzyme for a deep biophysical characterization, and to use it for the design of a new sensing device that could help the diagnosis of some neurological diseases at an early stage.

Research goals

The main goal of this project is the biophysical characterization of a recombinant D-serine dehydratase that binds D-serine by means of advanced techniques such as steady-state and time-resolved fluorescence spectroscopy and circular dichroism spectroscopy and the development of a optical biosensor for the detction of D-serine. It is of fundamental importance to characterize at molecular level a protein if biotechnological application are planned. In our project we plan to design an advanced fluorescence biosensor for monitoring the level of D-serine in serum and/or interstitial fluids by using as probe for the biosensor the D-serine dehydratase isolated from S. cerevisiae. In this context, our proposal is to joint the different and synergic expertise of the dr. D’Auria lab in biosensor design with the expertise of Dr. Turoverov in protein biophysics.

Last update: 20/04/2024