Research topics:
-Analytical separation techniques:
fundamentals, instrumentation and applications in the
biological and
biotechnological fields
-Development and application of
field-flow fractionation (FFF) techniques; foundamental
topics:
-
separation and characterization of macromolecules, colloids and
particulate of biological, field.
-limit of detection
-
hyphenation of FFF with chemiluminescence (CL) detection for high
sensitivity and high selectivity
analysis of dispersed bioanalytes.
-
instrumental development of miniaturized, disposable, biocompatible
FFF devices.
- optimization of channel composition for FFF separation in
bioanalytical field
Application research of FFF:
-
Invention of cell sorting methods for intact prokaryotic and
eukaryotic cells;
-
hyphenation of the developed cell sorting methods with CL, laser
scattering and mass spectrometric detection.
-
Invention of methods and device for stem cell
characterization/enrichment
-
Development of method for sample preparation (whole cells,
bacteria, protein and proteic complexes) from complex biological
matrices prior to specific analysis
- Development of nw format for flow-assisted immunoassay
-Development of a new hybrid fractionation method based on FFF and
biospecific reagents
Cell sorting: The
majority of cell sorting methodologies, including
Magnetic-Activated and Fluorescent-Activated Cell Sorting, make use
of immunological markers. For some applications, however, specific
antigenic markers may be lacking, or the labeling process itself
may affect viability or functionality of the sorted cells. New
techniques for label-free cell sorting would be, therefore, of
great interest particularly for applications to stem cells. In
collaboration with Prof. G.P. Bagnara (Dept. of Histology,
Embriology and Applied Biology, University of Bologna)
GrFFF-related techniques have been developed to sort adult human
stem cells. Such cells can be isolated from different tissues and
used in-vivo to regenerate damaged tissues, or ex-vivo to create
new tissues, with promising applications in regenerative medicine.
A new technique to enrich and sort multipotent human stem cells has
been invented and patented (WO07128737, owned by the University of
Bologna).
Moreover, different applications of FFF to
purify/enrich cells from clinical specimens were developed to
obtain cells with homogenous
properties to be used after the fractionation process.
Methodological a nd instrumental aspects for an esay integration in
more complex analysi system, were developed.
Immunological methods: in cooperation with the Group of Bioanalitical
Chemistry at the Dept. of Pharmaceutical Sciences (Prof. A. Roda)
new analytical formats are developed for flow-through immunometric
methods. Use of flow-through CL detectors online coupled with the
fractionation device (GrFFF-FIA-CL) allows to determine the analyte
concentration by comparing the fractographic peak area with a
proper response curve. Based on the intrinsic advantages of such an
approach, a non-competitive immunoassay has been developed, which
uses GrFFF-FIA-CL for the quantitation of whole cells (pathogenic
bacteria).
Analysis of dispersed
microparticles: In the framework
of the project "STARCHitechture" (University Strategic Projects -
Senior, coordinator P. Trost) FlFFF-MALS and gravitational FFF
(GrFFF) are applied to the separation and characterization of
starch granules and enzymatic degradation products from mutant
cereals.
