Piattaforma interdipartimentale di spettrometria di massa

Piattaforma interdipartimentale di spettrometria di massa UNIMIB
Logo Piattaforma interdipartimentale di spettrometria di massa UNIMIB

ENGLISH VERSION: https://www.btbs.unimib.it/en/platform for mass spectrometry

Una piattaforma di spettrometria di massa per l'analisi di piccole e grandi molecole da campioni purificati o da matrici complesse per quantificazione, caratterizzazione strutturale e studi “omici” con altissima risoluzione.

Strumentazione: Orbitrap Fusion™ Tribrid™ accoppiato a EASY-nLC 1000 UHPLC, TSQ Quantum™ Access MAX accoppiato a UltiMate 3000 UHPLC  (Thermo Fisher Scientific).

Responsabile: Prof. Rita Grandori

Pagina web: In costruzione!

Contatti: ms-center@unimib.it

Il laboratorio è dotato di due strumenti di Thermo Fisher Scientific, un Orbitrap Fusion e un TSQ Quantum Access MAX.

Orbitrap Fusion

Lo spettrometro di massa Orbitrap Fusion permette l’analisi di molecole e complessi supramolecolari nel range di 50-6000 m/z, con altissima risoluzione (fino a R=450.000), velocità di scansione (MSn fino a 20 Hz), accuratezza di massa (< 3 ppm), range dinamico (>  5.000) e sensibilità (100 fg di quantità totale di standard di reserpina).

Lo strumento è caratterizzato da una grande flessibilità di protocolli di scansione basati su modalità di frammentazione multiple, ossia la dissociazione indotta da collisione (CID), la dissociazione collisionale ad alta energia (HCD), la dissociazione da trasferimento di elettroni (ETD) e loro combinazioni, che possono essere eseguite in due celle di collisione differenti (quadrupolo o trappola ionica lineare) e analizzate con due analizzatori differenti (trappola ionica lineare o Orbitrap). Queste caratteristiche lo rendono adatto a studi di proteomica bottom-up, top-down e middle-down e permettono l’analisi ad elevati processività e dettaglio di piccole molecole, peptidi, proteine, modificazioni post-traduzionali, interazioni proteina-ligando, e altri polimeri.

La proteomica shotgun quantitativa può essere implementata con o senza l’utilizzo di isotopi stabili. Lo strumento è dotato di una sorgente per la ionizzazione elettrospray (ESI), sia standard che nano, e può essere accoppiato ad un UHPLC EASY-nLC 1000 a nanoflussi, permettendo l’analisi LC/MS automatizzata e performante di matrici biologiche complesse. L’analisi dei dati di proteomica è eseguita mediante il software Proteome Discoverer 2.2.

Servizi offerti:

  1. Preparazione del campione: sostituzione del tampone/desalinizzazione, digestione enzimatica in gel e in soluzione, arricchimento di peptidi, marcatura per quantificazione di proteine (iTRAQ e TMT).
  2. Analisi di spettrometria di massa: ESI-MS con infusione diretta o cromatografia liquida, proteomica shotgun con o senza marcatura, quantificazione mirata e mappatura di modificazioni post-traduzionali.
  3. Analisi dei dati e interpretazione: analisi di singoli spettri, ricerca in banca-dati, mappatura e quantificazione.

TSQ Quantum Access MAX

Lo strumento TSQ Quantum Access MAX è un analizzatore a triplo quadrupolo dotato di una sorgente di ionizzazione HESI e di una cella di collisione a 90° composta da un quadrupolo quadrato ad alta efficienza. Lo strumento permette l’analisi di molecole nel range di massa di 10-3000 m/z con velocità di scansione di 5000 µ/s e 25 ms per lo scambio tra modalità positiva e negativa. Le funzioni di scansione includono la scansione completa MS in Q1 o Q3, il monitoraggio di uno ione selezionato (SRM) in Q1 o Q3, il monitoraggio di reazioni selezionate (SRM), la scansione di ioni precursore, la scansione di ioni prodotto e la scansione della perdita del frammento neutro. In SRM è possibile definire fino a 3000 segmenti temporizzati e 2 ms di tempo di permanenza. Queste caratteristiche permettono l’analisi di centinaia di composti in una singola corsa. Inoltre, l’analisi MS/MS dato-dipendente e migliorata per la quantificazione (QED-MS/MS) fornisce simultaneamente la quantificazione e l’identificazione del composto.

Le applicazioni della quantificazione di piccole molecole spaziano dalla ricerca ambientale, farmaceutica e clinica all’ambito dell’alimentazione. Lo spettrometro di massa è accoppiato con un cromatografo liquido UltiMate LPG-3400SD che supporta 4 eluenti mediante una valvola dosatrice con flussi che possono essere impostati da 0,001 a 10 mL/min a pressioni fino a 620 bar. Questo sistema LC non solo supporta le applicazioni standard di cromatografia liquida, ma offre anche una completa compatibilità con separazioni veloci mediante UHPLC. L’acquisizione dei dati e l’analisi vengono eseguite con il software Xcalibur.

Servizi offerti:

  1. Preparazione del campione: estrazione (in fase solida, in fase solida dispersiva) e microestrazione (in fase liquida a singola goccia, liquido-liquido dispersiva, in fase solida, in fase solida dispersiva), marcatura per quantificazione
  2. Analisi di spettrometria di massa: ESI-MS con infusione diretta o cromatografia liquida
  3. Analisi dei dati e interpretazione: analisi di singoli spettri o cromatogrammi

Tariffario

ICMS.01A - In-gel protein Identification by bottom-up or middle-down MS - method development and analytical setting
ICMS.02A - In-gel protein Identification by bottom-up or middle-down MS - sample preparation and MS data collection: band excision, washing, reduction, alkylation, in-gel digestion (trypsin); peptides extraction and desalting; nUHPLC-MS/MS analysis (short gradient, HCD fragmentation, duplicate run)
ICMS.02B - ▼ In-gel protein Identification by bottom-up or middle-down MS - alternative/additional digestion protocol
ICMS.02C - In-gel protein Identification by bottom-up or middle-down MS - phosphopeptides-enrichment (TiO2)
ICMS.02D - In-gel protein Identification by bottom-up or middle-down MS - glycopeptides-enrichment (HILIC)
ICMS.02E - ▼ In-gel protein Identification by bottom-up or middle-down MS - alternative/additional enrichment protocol
ICMS.02F - In-gel protein Identification by bottom-up or middle-down MS - additional fragmentation (CID, ETD)
ICMS.03A - In-gel protein Identification by bottom-up or middle-down MS - data analysis: data processing; database search and protein identification; interpretation and report drafting
ICMS.03B - In-gel protein Identification by bottom-up or middle-down MS - PTM mapping
ICMS.04A - In-solution protein Identification by bottom-up or middle-down MS - method development and analytical setting
ICMS.05A - In-solution protein Identification by bottom-up or middle-down MS - sample preparation and MS data collection: in-solution reduction, alkylation, digestion (trypsin); peptides desalting; nUHPLC-MS/MS analysis (short gradient, HCD fragmentation, duplicate run)
ICMS.05B - ▼ In-solution protein Identification by bottom-up or middle-down MS - alternative/additional digestion protocol
ICMS.05C - In-solution protein Identification by bottom-up or middle-down MS - phosphopeptides-enrichment (TiO2)
ICMS.05D - In-solution protein Identification by bottom-up or middle-down MS - glycopeptides-enrichment (HILIC)
ICMS.05E - ▼ In-solution protein Identification by bottom-up or middle-down MS - alternative/additional enrichment protocol
ICMS.05F - In-solution protein Identification by bottom-up or middle-down MS - additional fragmentation (CID, ETD)
ICMS.06A - In-solution protein Identification by bottom-up or middle-down MS - data analysis: data processing; database search and protein identification; interpretation and report drafting
ICMS.06B - In-solution protein Identification by bottom-up or middle-down MS - PTM mapping
ICMS.07A - Qualitative proteomics by bottom-up or middle-down MS - method development and analytical setting
ICMS.08A - Qualitative proteomics by bottom-up or middle-down MS - sample preparation and MS data collection: in-solution reduction, alkylation, digestion (trypsin); peptides desalting; nUHPLC-MS/MS analysis (long gradient, HCD fragmentation, duplicate run)
ICMS.08B - Qualitative proteomics by bottom-up or middle-down MS - albumin and IgG immunodepletion (for biofluids)
ICMS.08C - ▼ Qualitative proteomics by bottom-up or middle-down MS - alternative/additional digestion protocol
ICMS.08D - Qualitative proteomics by bottom-up or middle-down MS - phosphopeptides-enrichment (TiO2)
ICMS.08E - Qualitative proteomics by bottom-up or middle-down MS - glycopeptides-enrichment (HILIC)
ICMS.08F - ▼ Qualitative proteomics by bottom-up or middle-down MS - alternative/additional enrichment protocol
ICMS.08G - Qualitative proteomics by bottom-up or middle-down MS - additional fragmentation (CID, ETD)
ICMS.09A - Qualitative proteomics by bottom-up or middle-down MS - data analysis: data processing; database search and protein identification; interpretation and report drafting
ICMS.09B - Qualitative proteomics by bottom-up or middle-down MS - PTM mapping
ICMS.10A - Label-free quantitative proteomics by bottom-up or middle-down MS - method development and analytical setting
ICMS.11A - Label-free quantitative proteomics by bottom-up or middle-down MS - sample preparation and MS data collection: in-solution reduction, alkylation, digestion (trypsin); peptides desalting; nUHPLC-MS/MS analysis (long gradient, HCD fragmentation, duplicate run)
ICMS.11B - Label-free quantitative proteomics by bottom-up or middle-down MS - albumin and IgG immunodepletion (for biofluids)
ICMS.11C - ▼ Label-free quantitative proteomics by bottom-up or middle-down MS - alternative/additional digestion protocol
ICMS.11D - Label-free quantitative proteomics by bottom-up or middle-down MS - phosphopeptides-enrichment (TiO2)
ICMS.11E - Label-free quantitative proteomics by bottom-up or middle-down MS - glycopeptides-enrichment (HILIC)
ICMS.11F - ▼ Label-free quantitative proteomics by bottom-up or middle-down MS - alternative/additional enrichment protocol
ICMS.11G - Label-free quantitative proteomics by bottom-up or middle-down MS - additional fragmentation (CID, ETD)
ICMS.12A - Label-free quantitative proteomics by bottom-up or middle-down MS - data analysis: data processing; database search and protein identifications; determination of relative normalized abundance; interpretation and report drafting
ICMS.12B - Label-free quantitative proteomics by bottom-up or middle-down MS - PTM mapping
ICMS.13A - Label-based quantitative proteomics by bottom-up or middle-down MS - method development and analytical setting
ICMS.14A - Label-based quantitative proteomics by bottom-up or middle-down MS - sample preparation and MS data collection: in-solution reduction, alkylation, digestion (trypsin); peptides desalting; nUHPLC-MS/MS analysis (long gradient, HCD fragmentation, duplicate run)
ICMS.14B - Label-based quantitative proteomics by bottom-up or middle-down MS - TMT labelling (up to 10 compared samples)
ICMS.14C - Label-based quantitative proteomics by bottom-up or middle-down MS - iTRAQ labelling (up to 4 compared samples)
ICMS.14D - Label-based quantitative proteomics by bottom-up or middle-down MS - albumin and IgG immunodepletion (for biofluids)
ICMS.14E - ▼ Label-based quantitative proteomics by bottom-up or middle-down MS - alternative/additional digestion protocol
ICMS.14F - Label-based quantitative proteomics by bottom-up or middle-down MS - phosphopeptides-enrichment (TiO2)
ICMS.14G - Label-based quantitative proteomics by bottom-up or middle-down MS - glycopeptides-enrichment (HILIC)
ICMS.14H - ▼ Label-based quantitative proteomics by bottom-up or middle-down MS - alternative/additional enrichment protocol
ICMS.14I - Label-based quantitative proteomics by bottom-up or middle-down MS - additional fragmentation (CID, ETD)
ICMS.15A - Label-based quantitative proteomics by bottom-up or middle-down MS - data analysis: data processing; database search and protein identifications; determination of relative normalized abundance; interpretation and report drafting
ICMS.15B - Label-based quantitative proteomics by bottom-up or middle-down MS - PTM mapping
ICMS.16A - Targeted quantitative proteomics by bottom-up MS - method development and analytical setting
ICMS.17A - Targeted quantitative proteomics by bottom-up MS - sample preparation and MS data collection: in-solution reduction, alkylation, digestion (trypsin); peptides desalting; targeted nUHPLC-MS/MS analysis (long gradient, HCD fragmentation, duplicate run)
ICMS.17B - Targeted quantitative proteomics by bottom-up MS - albumin and IgG immunodepletion (for biofluids)
ICMS.17C - ▼ Targeted quantitative proteomics by bottom-up MS - alternative/additional digestion protocol
ICMS.17D - Targeted quantitative proteomics by bottom-up MS - phosphopeptides-enrichment (TiO2)
ICMS.17E - Targeted quantitative proteomics by bottom-up MS - glycopeptides-enrichment (HILIC)
ICMS.17F - ▼ Targeted quantitative proteomics by bottom-up MS - alternative/additional enrichment protocol
ICMS.17G - Targeted quantitative proteomics by bottom-up MS - additional fragmentation (CID, ETD)
ICMS.18A - Targeted quantitative proteomics by bottom-up MS - data analysis: data processing; quantification of targeted proteins; interpretation and report drafting
ICMS.18B - Targeted quantitative proteomics by bottom-up MS - PTM mapping
ICMS.19A - ▼ Functional annotation of protemic results (networks, biological processes, pathways)
ICMS.20A - Native MS of proteins and protein complexes - method development and analytical setting
ICMS.21A - Native MS of proteins and protein complexes - sample preparation and MS data collection: protein desalting and buffer exchange; sample preparation before injection (dilution, denaturant and/or ligand addition etc.);  nano-ESI MS analysis (duplicate)
ICMS.21B - Native MS of proteins and protein complexes - protein-protein or protein-ligand dissociation by CID
ICMS.22A - Native MS of proteins and protein complexes - data analysis: data processing, mass deconvolution; conformational ensemble deconvolution by Gaussian fitting; estimation of solvent accessible surface area; interpretation and report drafting
ICMS.22B - Native MS of proteins and protein complexes - titration for folding or binding analysis
ICMS.23A - Proteoforms characterization by top-down MS - method development and analytical setting
ICMS.24A - Proteoforms characterization by top-down MS - sample preparation and MS data collection: desalting, buffer exchange, denaturation;  nano-ESI MS analysis (duplicate)
ICMS.24B - Proteoforms characterization by top-down MS - nano-ESI MS/MS analysis by HCD fragmentation (per proteoform)
ICMS.24C - Proteoforms characterization by top-down MS - nano-ESI MS/MS analysis by CID fragmentation (per proteoform)
ICMS.24D - Proteoforms characterization by top-down MS - supercharging agent addition; nano-ESI MS/MS analysis by ETD fragmentation (per proteoform)
ICMS.25A - Proteoforms characterization by top-down MS - data analysis: data processing, mass deconvolution; interpretation and report drafting
ICMS.25B - Proteoforms characterization by top-down MS - confirmation of protein identity from MS/MS spectra; PTMs identification and mapping
ICMS.25C - Proteoforms characterization by top-down MS - deconvolution of isobaric proteoform mixtures
ICMS.26A - Protein structural analysis by covalent labelling - method development and analytical setting
ICMS.27A - Protein structural analysis by covalent labelling - sample preparation and data collection: protein covalent modification (amino-groups, carboxyl-groups, etc.); in-solution reduction, alkylation, digestion (trypsin); peptides desalting; nUHPLC-MS/MS analysis (short gradient, HCD fragmentation, duplicate run)
ICMS.27B - ▼ Protein structural analysis by covalent labelling - alternative/additional digestion protocol
ICMS.27C - Protein structural analysis by covalent labelling - additional fragmentation (CID, ETD)
ICMS.28A - Protein structural analysis by covalent labelling - data analysis: data processing; determination of relative normalized abundance; interpretation and report drafting
ICMS.29A - Structural proteomics by limited proteolysis (LiP)-MS - method development and analytical setting
ICMS.30A - Structural proteomics by limited proteolysis (LiP)-MS - sample preparation and data collection: limited proteolysis (proteinase K) and complete digestion (trypsin); reference complete digestion (trypsin); peptides desalting
ICMS.30B - Structural proteomics by limited proteolysis (LiP)-MS - untargeted nUHPLC-MS/MS analysis on sample and reference (duplicate run)
ICMS.30C - Structural proteomics by limited proteolysis (LiP)-MS - targeted nUHPLC-MS/MS analysis on sample and reference (duplicate run)
ICMS.30D - ▼ Structural proteomics by limited proteolysis (LiP)-MS - alternative/additional digestion protocol
ICMS.31A - Structural proteomics by limited proteolysis (LiP)-MS - data analysis: data processing; determination of proteolytic sites and differential analysis; interpretation and report drafting
ICMS.32A - H/D exchange analysis on intact protein - method development and analytical setting
ICMS.33A - H/D exchange analysis on intact protein - sample preparation and data collection: protein desalting and lyophilization; sample preparation for isotopic labelling (exchange-in or exchange-out)
ICMS.33B - H/D exchange analysis on intact protein - nano-ESI MS analysis in real time
ICMS.33C - H/D exchange analysis on intact protein - aliquot sampling, quenching and nano-ESI MS analysis
ICMS.33D - H/D exchange analysis on intact protein -  ETD fragmentation on selected masses & charge states
ICMS.34A - H/D exchange analysis on intact protein - data analysis: data processing; interpretation and report drafting
ICMS.34B - H/D exchange analysis on intact protein - analysis of protein conformational dynamics
ICMS.34C - H/D exchange analysis on intact protein - solvent-exposed region mapping
ICMS.35A - Small molecules analysis from purified samples - method development and analytical setting
ICMS.36A - Small molecules analysis from purified samples - sample preparation for direct injection
ICMS.36B - Small molecules analysis from purified samples - low resolution ESI-MS analysis (TSQ Quantum Access Max)
ICMS.36C - Small molecules analysis from purified samples - high resolution ESI-MS analysis (Orbitrap Fusion)
ICMS.36D - Small molecules analysis from purified samples - ESI-MS/MS analysis by CID fragmentation (TSQ Quantum Access Max)
ICMS.36E - Small molecules analysis from purified samples - ESI-MS/MS analysis by CID fragmentation (Orbitrap Fusion)
ICMS.36F - Small molecules analysis from purified samples - additional ESI-MS/MS analysis by HCD/ETD fragmentation (Orbitrap Fusion)
ICMS.37A - Small molecules analysis from purified samples - data analysis: data processing, mass deconvolution
ICMS.37B - Small molecules analysis from purified samples - interpretation of MS/SM spectra and report drafting
ICMS.38A - Targeted small molecules quantitation in complex mixtures - method development and analytical setting (15 determinations included)
ICMS.39A - Targeted small molecules quantitation in complex mixtures - single drop microextraction
ICMS.39B - Targeted small molecules quantitation in complex mixtures - dispersive liquid-liquid microextraction
ICMS.39C - Targeted small molecules quantitation in complex mixtures - solid-phase (micro)extraction
ICMS.39D - Targeted small molecules quantitation in complex mixtures - dispersive solid-phase (micro)extraction
ICMS.39E - Targeted small molecules quantitation in complex mixtures - HPLC-MS/MS, standard gradient
ICMS.39F - Targeted small molecules quantitation in complex mixtures - UHPLC-MS/MS, standard gradient
ICMS.39G - Targeted small molecules quantitation in complex mixtures -labelling for unknowns quantitation
ICMS.40A - Targeted small molecules quantitation in complex mixtures - data analysis: data processing; quantitation of targeted molecules; interpretation and report drafting

Inoltre, offriamo speciali pacchetti di analisi ad un prezzo scontato, come lo sviluppo di metodi di quantificazione personalizzati.

Sconti speciali verranno applicati per campioni o analisi numerose. Scarica il nostro Listino [Brochure] e contattaci per maggiori informazioni!