Multi - user equipment approved in grant 2012/50191-4: mass spectrometer impact II - Bruker

Abstract

We have been using mass spectrometry since 1994 as a control tool in the parallel synthesis of peptides established by us since 1990 and this experience has accredited us to use mass spectrometry during development of new projects, as well as in the complementary projects. For 03 years UNIFESP has been structuring the area of oncology, adding researchers from different areas and different institutions of the State of São Paulo, with whom we already had collaborations on account of the peptides and now we are able to contribute with mass spectrometry. We are currently involved in collaborations with Dr. André Vettore and Dr. Giselle Zanker, professors from UNIFESP-Campus Diadema in the identification of classes of proteases of head and neck tumor cells; of brain and leukemic cells using substrates and inhibitors. The next step will be the identification of the proteins present in the cells, as well as the absolute and relative quantification of these proteins. Other projects are being developed aiming at the use of this tool, mainly in the field of clinical oncology for various purposes, such as biomarkers, being used for differentiation of the plasma secret in peptidoma and identification of post-translational modifications of patients with gastric cancer. This project will be performed in collaboration with Dr. Nora Manouikian Forones, Professor of the EPM/UNIFESP, with emphasis on gastroenterology/cancerology. From this thematic project (# 12 / 50191-4), a Nano-HPLC (Thermo) was purchased to be attached to an automatic dispenser on MALDI-TO / TOF (Proteineer FC, Bruker) plates and a Workstation with programs (Peaks studio, Waterloo, Canada; Profile Analysis and Protein Scape, Bruker Daltonics, Bremen, Germany) for proteomic analyzes. However, this system does not allow carrying out the experiments exposed above. The objective of this request is to make possible the purchase of the mass spectrometer Impact II (Bruker), already quoted by the company responsible, from the financial resource available in the project without budget increase, since the company manufacturer of the equipment already made feasible the sale according to the available resource. The equipment will be multiuser, since there are already collaborations with several researchers in the areas of basic and clinical research of the Escola Paulista de Medicina, as well as researchers from UNIFESP Campus - Diadema / SP, who they are looking forward to exploring applications that this tool can provide. The nanoLC system coupled to mass spectrometer and softwares, can perform the identification of proteins (proteomics topdown and botton-up), sequencing and identification of novel compounds, peptidomic and metabolomic analyze, besides to perform additional operations, such as label-free quantification, since it is possible to evaluate the chromatogram and intensity / area of the precursor ions and all tools are compatible with date acquired. Demands in the area of metabolomics may also be used since our HPLCs are compatible with the system. Finally, it is important to emphasize that the infrastructure installed on account of this thematic project, already mentioned above and added with the acquisition of equipment will be part of an ample project of metabolic analysis in the rejection of kidneys transplanted by Hospital Rim, which 900 to 950 transplants/year, being the most active kidney transplant center in the world. In this project, we will use the mass spectrometry associated to Magnetic Nuclear Resonance (NMR) in metabolomics analysis whose equipment and infrastructure have just been installed in INFAR under the coordination of Dra. Helena Nader. In addition, Bruker company, also manufacturer of NMR, will provide technical and logistic support for this technological innovation of MS / RMN association. (AU)