ADOR - Hydro-enzyme (DCM.AD006.411)
Thematic area
Chemical sciences and materials technology
Project area
Chimica e tecnologia dei materiali (DCM.AD006)Structure responsible for the research project
Istituto di Scienze e Tecnologie Chimiche "Giulio Natta" (SCITEC)
Project manager
MARCELLA CHIARI
Phone number: 0228500035
Email: marcella.chiari@scitec.cnr.it
Abstract
The term hydrogel defines a viscoelastic solid-like deformable material composed of a dispersion of a substantially diluted cross-linked polymeric network in a continuous aqueous medium. Its ability to maintain the shape while absorbing a large amount of water or aqueous media and the high permeability towards small molecules, metabolites, and other water-soluble components makes the hydrogel a suitable microenvironment that increases the stability of macromolecules while maintaining their native conformation.
The features of hydrogels have been widely exploited in enzyme-based biosensors. The project aims to develop a novel click-chemistry-based printable hydrogel for DNA microarray with application in biosensing. Click-chemistry reactions are renowned for their high reactivity, selectivity, and mild conditions. These characteristics make them suitable for producing hydrogels with micro three-dimensional structures that can accommodate different proteins, particularly enzymes while maintaining their active conformations and catalytic activity. The project aims to explore CRISPR-Cas12a, an RNA-guided enzyme that binds and cuts DNA, in a multiplex platform for parallel detection of
Goals
The first objective of the project is the development of a suitable click-hydrogel that can be printed in a microarray format on a solid support. Among the different click-chemistry reactions that could be employed for this purpose, Cu-ACC and SPAAC reactions present several advantageous features that could simplify the printing procedure of the hydrogel. Both have a fast reaction kinetics, occur at room temperature and are not affected by solution parameters as pH or ionic strength. Furthermore, azide and alkyne residues have limited cross reaction with biological targets.
The second objective is to optimize a printing strategy using a non-contact piezoelectric spotter that allows depositing picolitres of solution on a solid support that could be glass, silicon, or plastic in different formats. Immobilization of bio-receptors on a hydrogel surface is one of the most critical steps in biosensor design.
The third objective is to demonstrate the diagnostic potential of the proposed biosensing platform, by developing a DNA sensor based on the use CRISPR-Cas12a enzyme
Start date of activity
01/01/2021
Keywords
HYDROGEL, ENZYME, MICROARRAY
Last update: 07/06/2025