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Università Vita-Salute San Raffaele

3 ARTICLES PUBLISHED IN JoVE

Medicine

From a 2DE-Gel Spot to Protein Function: Lesson Learned From HS1 in Chronic Lymphocytic Leukemia

Benedetta Apollonio^1,2, Maria Teresa Sabrina Bertilaccio¹, Umberto Restuccia³, Pamela Ranghetti¹, Federica Barbaglio¹, Paolo Ghia^1,4, Federico Caligaris-Cappio^1,4, Cristina Scielzo¹

¹Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute, ²Department of Haemato-Oncology, King's College London, ³IFOM, FIRC Institute of Molecular Oncology, ⁴Università Vita-Salute San Raffaele

Here we describe a protocol that couples two proteomic techniques, namely 2-dimensional Electrophoresis (2DE) and Mass Spectrometry (MS), to identify differentially expressed/post-translational modified proteins among two or more groups of primary samples. This approach, together with functional experiments, allows the identification and characterization of prognostic markers/therapeutic targets.

Cancer Research

Immunoglobulin Gene Sequence Analysis In Chronic Lymphocytic Leukemia: From Patient Material To Sequence Interpretation

Andreas Agathangelidis^*1, Lesley Ann Sutton^*2,3, Anastasia Hadzidimitriou¹, Cristina Tresoldi⁴, Anton W. Langerak⁵, Chrysoula Belessi⁶, Frederic Davi⁷, Richard Rosenquist^2,3, Kostas Stamatopoulos^1,2, Paolo Ghia⁸

¹Institute of Applied Biosciences, Centre for Research and Technology Hellas, ²Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, ³Department of Molecular Medicine and Surgery, Karolinska Institutet, ⁴Division of Immunology, Transplantation and Infectious, IRCCS San Raffaele Scientific Institute, ⁵Department of Immunology, Laboratory for Medical Immunology, Erasmus University Medical Center, ⁶Hematology Department, Nikea General Hospital, ⁷Assistance publique - Hôpitaux de Paris (AP-HP), Hopital Pitié-Salpêtrière, Department of Hematology, and UPMC University Paris 06, UMRS 1138, ⁸Division of Experimental Oncology, IRCCS Istituto Scientifico San Raffaele and Università Vita-Salute San Raffaele

Herein, we present a protocol that details the technical aspects and essential requirements to ensure robust IG gene sequence analysis in patients with chronic lymphocytic leukemia (CLL), based on the accumulated experience of the European Research initiative on CLL (ERIC).

Biology

Setting a Successful Sorting for Extracellular Vesicle Isolation

Monica Romanò^*1, Daniela Boselli^*1, Enrico Ragni², Laura de Girolamo², Chiara Villa^1,3

¹FRACTAL - Flow cytometry Resource, Advanced Cytometry Technical Applications Laboratory, IRCCS Ospedale San Raffaele, ²Laboratorio di Biotecnologie Applicate all'Ortopedia, IRCCS Istituto Ortopedico Galeazzi, ³Università Vita-Salute San Raffaele

This protocol provides a detailed description of sorting extracellular vesicles (EVs) released by mesenchymal stromal cells. In particular, it focuses on the instrument setting and the optimization of the sorting conditions. The goal is to sort extracellular vesicles while preserving their characteristics.