Separative Techniques and Affinity Interactions Lab
Macroarea: Pharmaceutical Analysis
ERC Sectors: PE4_5 Analytical Chemistry, PE5_6 New materials: oxides, alloys, composite, organic-inorganic hybrid, nanoparticles, PE11_9 Nanomaterials engineering, e.g. nanoparticles, nanoporous materials, 1D & 2D nanomaterials, PE5_18 Medicinal chemistry, LS7_3 Nanomedicine, LS7_2 Medical technologies and tools (including genetic tools and biomarkers) for prevention, diagnosis, monitoring and treatment of diseases
Lab Managers: Ersilia De Lorenzi (Full Professor), Massimo Serra (Technician)
Junior Staff: Cecilia Contardi (PhD student)
1. CHARACTERIZATION OF MOLECULARLY IMPRINTED NANOPARTICLES. Molecularly Imprinted polymers (MIPs) are designed in-house to detect and sense previously inaccessible tumor markers or to discover novel disease biomarkers. MIPs are characterized during their design and development for final material optimization and reproducibility. Characterization of MIPs in different formats, including nanoparticles is carried out by capillary electrophoresis (CE), liquid chromatography (LC), dynamic light scattering, zeta potential, atomic force microscopy. Template-MIP interaction studies are performed by affinity CE (dynamic complexation CE, frontal CE) and frontal LC.
2. ANALYSIS AND CHARACTERIZATION OF PROTEINS AS DRUG TARGETS Advanced analytical and spectroscopic techniques for separation, identification, characterization, quantification and evaluation of proteins as drug targets (charge variants, folding conformers and/or aggregates of proteins with pathological or therapeutic potential). In particular studies are carried out on amyloidogenic proteins.
3. FROM AFFINITY STUDIES TO DRUG DISCOVERY. Integrated strategies to correlate aggregation state, structure and toxicity of Aß 1-42 oligomers as drug targets (capillary electrophoresis, ATR-FTIR, TEM, in silico simulations, biological tests on cell lines). Evaluation of novel curcumin analogues as a potential therapeutic agents against Alzheimer’s Disease. Anti-oligomeric, anti-fibrillogenic, anti-inflammatory, anti-oxidant activity are investigated on cell lines, primary neuronal cells or peripheral blood mononuclear cells from AD patients.
- Groves, K. et al: REFERENCE PROTOCOL TO ASSESS ANALYTICAL PERFORMANCE OF HIGHER ORDER STRUCTURAL ANALYSIS MEASUREMENTS: RESULTS FROM AN INTERLABORATORY COMPARISON (2021) Analytical Chemistry, 93(26), pp. 9041-9048
- Bisceglia, F. et al: PRENYLATED CURCUMIN ANALOGUES AS MULTIPOTENT TOOLS TO TACKLE ALZHEIMER'S DISEASE (2019) ACS Chemical Neuroscience, ACS Chemical Neuroscience, 10(3), pp. 1420-1433
- Bisceglia, F. et al: AN INTEGRATED STRATEGY TO CORRELATE AGGREGATION STATE, STRUCTURE AND TOXICITY OF Aß 1–42 OLIGOMERS (2018) Talanta, 188, pp. 17-26
- De Lorenzi, E. et al: EVIDENCE THAT THE HUMAN INNATE IMMUNE PEPTIDE LL-37 MAY BE A BINDING PARTNER OF AMYLOID-Β AND INHIBITOR OF FIBRIL ASSEMBLY (2017) Journal of Alzheimer's Disease, 59(4), pp. 1213-122
- Bertoletti, L. et al: EVALUATION OF CAPILLARY ELECTROPHORESIS-MASS SPECTROMETRY FOR THE ANALYSIS OF THE CONFORMATIONAL HETEROGENEITY OF INTACT PROTEINS USING BETA2-MICROGLOBULIN AS MODEL COMPOUND (2016) Analytica Chimica Acta, 945, pp. 102-109
- Bertoletti, L. et al: CAPILLARY ELECTROPHORESIS ANALYSIS OF DIFFERENT VARIANTS OF THE AMYLOIDOGENIC PROTEIN Β2-MICROGLOBULIN AS A SIMPLE TOOL FOR MISFOLDING AND STABILITY STUDIES (2015) Electrophoresis, 36(19), pp. 2465-2472
- Bertoletti, L. et al: ADVANCED GLYCATION END PRODUCTS OF BETA2-MICROGLOBULIN IN UREMIC PATIENTS AS DETERMINED BY HIGH RESOLUTION MASS SPECTROMETRY (2014) Journal of Pharmaceutical and Biomedical Analysis, 91, pp. 193-201
- Brogi, S. et al: DISEASE-MODIFYING ANTI-ALZHEIMER'S DRUGS: INHIBITORS OF HUMAN CHOLINESTERASES INTERFERING WITH Β-AMYLOID AGGREGATION (2014) CNS Neuroscience and Therapeutics, 20(7), pp. 624-632
- Butini, S. et al.: MULTIFUNCTIONAL CHOLINESTERASE AND AMYLOID BETA FIBRILLIZATION MODULATORS. SYNTHESIS AND BIOLOGICAL INVESTIGATION (2013) ACS Medicinal Chemistry Letters, 4(12), pp. 1178-1182
- Bertoletti,L. et al.: SEPARATION AND CHARACTERISATION OF BETA2-MICROGLOBULIN FOLDING CONFORMERS BY ION-EXCHANGE LIQUID CHROMATOGRAPHY AND ION-EXCHANGE LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY (2013) Analytica Chimica Acta, 771, pp. 108-114
National collaborations
ACADEMIC INSTITUTIONS
Bice Conti, Rossella Dorati, Cristina Lanni, Department of Drug Sciences, University of Pavia
Maddalena Patrini, Department of Physics, University of Pavia;
Morena Zusso, Department of Pharmaceutical and Pharmacological Sciences,University of Padova
Federica Belluti, Department of Pharmacy and Biotechnology, University of Bologna;
Giulio Vistoli, Department of Pharmaceutical Sciences,University of Milano Statale
Sofia Giorgetti, Department of Molecular Medicine, University of Pavia
Antonino Natalello, Department of Biotechnology and Biosciences, University of Milano Bicocca
Laura Verga, IRCCS Policlinico San Matteo, Pavia Marcella Chiari, Institute of Chemistry of Molecular Recognition, CNR, Milano
Carlo Morasso, Istituto Neurologico Mondino, Pavia
INDUSTRY
Indena SpA, Settala (MI)
Merck-Serono, Guidonia (RM)
International collaborations
ACADEMIC INSTITUTIONS
Börje Sellergren, University of Malmö, Sweden, Ian Nicholls, University of Linnaeus, Sweden
Knut Rurack, Federal Institute for Materials Research and Testing
Panagiotis Manesiotis, Queen’s University, Belfast, United Kingdom
Annelise Barron, Stanford University, Stanford, CA, USA
INDUSTRY
LGC Ltd.,Teddington, UK
Marie Skłodowska-Curie Actions (H2020-MSCA-ITN-2016, 722171-Biocapture)
Competences
Development and validation of analytical methods for small molecules and proteins by capillary electrophoresis (CE-UV), including charge variant profiles of antibodies and pegylated proteins. Feasibility studies and CE method optimization for daily use on drug substance and drug product. Studies on drug-drug, drug-protein, drug-nanoparticle interactions in solution by affinity CE; Analysis of nanoparticles by CE. Development and validation of analytical methods for small molecules and proteins by HPLC-UV, HPLC-DAD. Feasibility studies and HPLC method optimization for daily use on drug substance and drug product.
Instrumentation
n. 2 CE (Agilent and Beckman) with DAD detector; n.2 HPLC (Agilent) with DAD detector; -n. 1 HPLC (Agilent) with UV-Vis detector