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Acquity MClass UPLC + Synapt G2-Si for Proteomics

Prof. Daniel Martins-de-Souza, PhD

University of Campinas (UNICAMP)

FAPESP Process: 2014/10068-4

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Contents

Where is this MUE?

The Lab of Neuroproteomics

Acquity MClass UPLC + Synapt G2-Si

Ion mobility

Timeline of equipment installation

Who is using this MUE?

Obtained results

Publications

University of Campinas (UNICAMP)

facebook.com/neuroproteomics

facebook.com/neuroproteomics

facebook.com/neuroproteomics

Department of Biochemistry, Institute of Biology, UNICAMP

Acquity MClass UPLC + Synapt G2-Si

Acquity MClass UPLC + Synapt G2-Si

2D-Ultraperformance Liquid Chromatography + Q-TOF

State-of-the-art quantitative proteomics

MSE

(data independent analysis)

State-of-the-art quantitative proteomics

Ion Mobility

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Proteome quantitation based in four

dimensions

• Retention Time

• Mass/Charge ratio

• Peak Intensity

• Drift Time

UDMSE (ultra definition) (Nat Methods. 2014 Feb;11(2):167-70.)

Label-free proteomics

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MUE´s Installation

Timeline

Laboratory of Neuroproteomics

UNICAMP

Young Investigator Award: (FAPESP Process: 2013/08711-3)

01.Feb.2014

MUE Award: (FAPESP Process: 2014/10068-4)01.Sep.2014

Equipment arrived in the lab: 10.Nov.2014

Lab Physical and Electrical Refurbishment (!!)30.Apr.2015

Equipment Installation: 18.May 2015 to 10.Jun.2015

First Samples Run: 15.Jun.2015

(Hardware and software tests)

Training: 03 to 07.Aug.2015

Equipment ready for use by capacitated people: 10.Aug.2015

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Lab RefurbishmentStarted in Oct. 1st, 2014

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Lab RefurbishmentStarted in Oct. 1st, 2014

+ nanoAcquity UPLC +

Synapt G2-Si

arrived at UNICAMPNov. 10th, 2014

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nanoAcquity UPLC + Synapt G2-Si

at UNICAMPMay, 2015

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InstallationStarted in May. 18th, 2015

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InstallationJun, 2015

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TrainingAug, 2015

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CurrentlyJun. 10th, 2016

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Mass Spectrometry Lab2016

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MUE

Users' committee

13 researchers of 6 different

departments from UNICAMP,

USP and AC Camargo

8 researchers (*) have already

had access to the equipment

1) UNICAMP (3 institutes)

a) Dept of Biochemistry and Tissue Biology, IB

Eneida de Paula (Head of Department)

Sérgio Marangoni (*)

Carmen V. Ferreira

Claudio C. Werneck (*)

b) Dept of Genetics, IB

Gonçalo G. Pereira (*)

Leonilda M. Santos (*)

Alessandro S. Farias (*)

Fábio T. Costa

c) Dept of Organic Chemistry, IQ

Fábio C. Gozzo (*)

d) Dept of Medical Genetics, FCM

Íscia L. Cendes (*)

2) USP

a) Institute of Psychiatry, FMUSP

Wagner F. Gattaz (*)

Helena P. Brentani

3) A.C. Camargo Cancer Center

Lab of Medical Genetics

Emmanuel Dias-Neto (*)

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MUE Availability

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MUE Availability

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MUE Availability

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Laboratory of Neuroproteomics

UNICAMP

Equipment ready for use by capacitated people: 10.Aug.2015

Up to 10.Jul.2016 = 11 months

31 different groups have used the instrument

MUE´s Installation

Timeline

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Groups per Country

Germany:

LMU and Uni Magdeburg

United Kingdom:

University of Cambridge

Australia:

James Cook University

United States:

University of Texas

Japan:

University of Fukui

Brazil25

Germany2

United Kingdom

1

Australia1

United States 1 Japan

1

+Users Per State,

in Brazil

Rio de Janeiro:

UFRJ/IDORProf. Stevens K. Rehen

Waters Co.Dr. Gustavo Souza

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Ceará: UFCProf. Claudia Pessoa

Rio Grande do Sul:UFRGSProf. Fábio Klamt

Minas Gerais: UFMGProf. Breno Satler Diniz

São Paulo20

Rio de Janeiro

2

Ceará1

Rio Grandedo Sul

1

Minas Gerais1

+Users in São Paulo

USP-SP:Prof. Wagner F. Gattaz

USP-RP:Prof. Vitor M. FaçaProf. Fernando Barbosa Jr.

AC Camargo:Dr. Emmanuel Dias-NetoDr. Vilma Regina Martins

UNESP-BotucatuProf. José Antônio Dell´Áqua Jr.

Unifesp – EPMProf. Mirian Hayashi

Structural Genomics Consortium (SGC)Dr. Opher Gileadi

INCORProf. Francisco Martins Laurindo

UNICAMP10

USP-SP; 2

USP-RP; 2

AC Camargo; 2

UNESP; 1

Unifesp; 1

SGC; 1

INCOR; 1

+Users at UNICAMP

Institute of Biology:

Prof. Jorg Kobarg

Prof. Sergio Marangoni

Prof. Fernanda R. Gadelha

Prof. André DamásioProf. Henrique Marques-Souza

Institute of Chemistry:

Prof. Fabio C. Gozzo

Prof. Marcos N. Eberlin

Prof. Alessandra Sussulini

Faculty of Medical Sciences:

Prof. Íscia Lopes-Cendes

Dr. Alexandre Nowill

IB5

IQ3

FCM2

+Machine time

Major use:

Lab of Neuroproteomics

In 2015: 56%

In 2016: 40%

UNICAMP:

66% (78%) of the time

São Paulo:

79% (91%) of the time

LNP48%

UNICAMP18%

São Paulo13%

Other States

5%

Foreigns4%

Maintenance12%

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Machine timededicated to

collaborators

São Paulo:

59% (82%) of the time

UNICAMP34%

São Paulo25%

Other States10%

Foreigns8%

Maintenance

23%

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ResultsAcquity MClass UPLC + Synapt G2-Si for Proteomics

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Collaborative work

Dr. Vilma R. Martins(AC Camargo)

Chemotherapy-resistant gastric cancer

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Collaborative work

Prof. Francisco M. Laurindo(InCor, SP)

Protein disulfide isomerase and cardiovascular biomarkers

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Garcez et al. 2016. PeerJ Preprints 4:e2033v1 Collaborative work/Publications

Prof. Stevens K. Rehen(IDOR/UFRJ)

Zika-infected human neurospheres

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Collaborative work/Publications

Dr. Fábio Passetti(FioCruz, RJ)

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LNP Publications

Veronica Saia-Cereda

(MSc)

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LNP Publications

Published 9 months ago

Veronica Saia-Cereda

(MSc)

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LNP Publications

Signaling

pathways

(ephrin B)

Astrocytes

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Proteomic Technology

Dr. Juliana S. Cassoli(Lab of Neuroproteomics, UNICAMP)

Comprehensive identification proteins

expressed by human oligodendroglia cells

FAPESP: 2014/14881-1

223,531 ion peptides and

10,614 proteinswere identified

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Proteomic Technology

Dr. Juliana S. Cassoli(Lab of Neuroproteomics, UNICAMP)

Label-free quantification of

human oligodendroglia cells proteins

FAPESP: 2014/14881-1

Immature x Mature Oligodendrocytes

493 proteinswere differentially expressed

96.6% overlap!

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Biomarker research

Sheila Garcia(PhD Student)

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+ aspects

- aspects

Multiuser Equipment

Promote collaboration

People capacitation

Optimize equipment investment

Not facility

“Just need some data to a grant report”

Consumables and maintenance ($)

Equipment time for the host group

Dr. Juliana S. Cassoli

Dr. Juliana M. Nascimento

Dr. Adriano Aquino

Paulo A. Baldasso (TA, BSc)

Sheila Garcia (PhD Student)

Verônica Saia-Cereda (PhD student)

Aline G. Santana (MSc student)

Caroline B Teles (MSc student)

Giuliana Zuccoli (MSc student)

Danielle Junqueira (BSc student)

Guilherme Reis (BSc student)

Dayene Delgado (BSc student)

Lícia Costa (BSc student)

FAPESP Process 13/08711-3Developing a predictive test for a successful medication response and understanding

the molecular bases of schizophrenia through proteomics

FAPESP Process 14/10068-4

Multi-user equipment for proteomics

Funding

CNPq project: 460289/2014-4

Phosphoproteome alteration in schizophrenia

brains

FAEPEX project: 0665-15/2015

Proteome analysis of schizophrenia