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Pierluigi Reschiglian

Full Professor

Department of Chemistry "Giacomo Ciamician"

Academic discipline: CHIM/01 Analytical Chemistry


Development and application of field-flow fractionation (FFF) techniques.

Fundamental research in FFF concerns:
(i) separation and characterization of macromolecules, colloids and particulate of biological, environmental and alimentary field.
(ii) characterization of nano- and microdispersed analytes by coupling FFF with uncorrelated techniques for morphological analysis.
(iii) hyphenation of FFF with chemiluminescence (CL) detection for high sensitivity and high selectivity analysis of dispersed bioanalytes. Development of flow assisted immunoassays and biosensors.
(iv) instrumental development of miniaturized, disposable, biocompatible FFF devices.

Application research topics of FFF:
i) Invention of cell sorting methods for prokaryotic and eukaryotic cells;
ii) hyphenation of the developed cell sorting methods with CL, laser scattering and mass spectrometric detection.

iii) characterization of protein expression of cells and native protein by coupling hollow fiber field flow fractionation and mass spectrometry for microorganism protein profiling, study of functional and allosteric proteins, characterization of recombinant proteins of pharmaceutical interest.

Analysis of nanoparticles: Synthesis and characterization of nanomaterials are the basis of future developments and applications of nanotechnologies, from electronics to biomedical technologies. The ability of hydrodynamic field FFF (FlFFF) for the analysis of nanodispersed analytes has been investigated in the fields of material science for nano-biotech applications. FlFFF has been, for instance, applied to the fractionation and size analysis of silica and titanium dioxide synthesized in the presence of fluorescent modifiers.  FlFFF showed able to control the synthesis of such multi-chromophoric structures, which have been then further characterized by using spectrometric and microscopic techniques. FlFFF has been also applied to fractionate functionalized carbon nanotubes (CNT). The project "LIGHT4HEALTH" (University Strategic Project - Junior, coordinator N. Zaccheroni) involves application of FlFFF coupled with multiangle laser scattering (MALS) detection for the analysis of fluorescent nanoparticles for applications in medicine.

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.

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).

Protein analysis: In a formal cooperation agreement with Wyatt Technology Europe (Germany) FlFFF-MALS has been developed from an instrumental and methodological point of view for the analysis and characterization of serum lipoproteins. Amyloidoses are a class of human diseases that arise from protein misfolding and subsequent aggregation into amyloid fibrils. In Alzheimer's disease (AD), a key role is played by a nucleation-dependent self-aggregation of the A β 1-42 peptide. . In cooperation with Prof. E. De Lorenzi (Dept. of Pharmaceutical Chemistry, University of Pavia), FlFFF-MALS has been applied to monitor the Aβ1-42 aggregation process through the separation of transient species and a broad-range, mass/size characterization (~ 5-1500 nm) of the prefibrillar and fibrillar Aβ1-42 aggregates. The original hyphenation of mass spectrometry and hollow-fiber (HF) FlFFF has been also developed for the analysis of intact proteins and protein complexes in native form. Functional recombinant protein drugs and complex protein samples such as whole human blood serum have been characterized.

In the framework of the locally coordinated project PRIN 2006 (Integration of novel separation and mass spectrometry technologies for third-generation proteomics, National coordinator Prof. G. Marino) , and of the Significant Research Project within the Bilateral Cooperation Protocol Italy-Korea 2007-2009, HF FlFFF and FlFFF-MALS are coupled with shot-gun proteomics techniques such as (nano/chip)LC-ESI(Ion-Trap)-MS/MS for the identification of native protein complexes in serum and sub-cellular fractions.

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).

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