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Dr. Pradip Chaudhari
Officer In Charge: Dr. Pradip Chaudhari

This facility focusses on preclinical animal imaging and research on radiopharmaceuticals. The diagnostic radionuclides - technetium-99m and fluorine-18 complexes are being evaluated for their utility in imaging and monitoring cancer xenografts in mouse models. In 2017, the facility supported 17 preclinical PET, SPECT and CT imaging studies on rodents for basic and translational research projects from ACTREC, other DAE units, academic institutes and pharmaceutical industries.

A majority of the studies were proof-of-concept studies, normal tracer uptake studies and in vivo tumor uptake studies - some involving validation of liver and brain xenograft and orthotopic models. The facility also designs imaging protocols for ex vivo bone imaging and analysis utilizing high resolution microCT, development of animal models, data quantitation, and analysis. During 2017, the facility conducted a major imaging study for the pharmaceutical industry. The facility also runs a complete cancer care program for the diagnosis and treatment of pet animals suffering from spontaneous cancer through its animal oncology clinic. During 2017, 122 referral cases underwent major/ minor surgeries, single or combination drug chemotherapy, radiation therapy or a combination of these, based on clinical requirement.

The animal cancer biorepository maintains biological material (blood, fresh-frozen/ formalin-fixed/ FFPE tissues) obtained during diagnosis and treatment, and utilizes them for research on comparative aspects of animal and human cancers. The senior faculty organized a CME on ‘Preclinical imaging and drug discovery’ at ACTREC from 20th to 22nd September 2017. The facility accepted six trainees during the year – one for Bachelor’s and four for Master’s dissertation, one for research experience, and one observer. Orientation tours of the facility were conducted during the year for visiting students and other visitors.

Dr Rajiv Sarin
Dr Rajiv Sarin

The Next Generation Sequencing (NGS) facility at ACTREC has a HiSeq 1500 from Illumina and an Ion Torrent Personal Genome Machine from Thermofisher. The machines were used by in house scientists as well as those from BARC and IISER, Pune during 2017.

Transcriptome analysis was carried out on the Hiseq 1500 to understand the expression of genes in oral cancer cell lines, in human samples exposed to low dose radiation, and in medullary thyroid cancer.

Exome sequencing was carried out to profile mutations in individuals exposed to radiation, for de novo sequencing of an insect exome, and to find differences in the penetrance and disease aggression of germline mutations in inherited cancers. After DNA/ RNA sample quality checks, exome and transcriptome libraries for 55 samples were prepared in house and sequenced on the HiSeq 1500. Good quality data was obtained and downstream bioinformatics analysis was done with open resource tools by the users.

Ion Torrent PGM libraries for targeted resequencing of BRCA1 and BRCA2 genes were prepared in 108 cases of hereditary breast and ovarian cancer syndrome. Pathogenic mutations were identified in 45 families.

MSF
Facility In Charge: Mr. Shashadhar Dolas

Overview

The Mass Spectrometry facility at ACTREC houses four state-of-the-art mass spectrometry platforms connected to high performance liquid chromatography systems – (1) Orbitrap Exploris 480, Thermo, (2) ESI/MALDI Q-TOF Model Synapt XS (3) Triple Quadrupole Model Xevo TQ Absolute and (4) The Nano-LC (ABSCIEX, Eksigent)-ESI-Q-TOF Model Triple TOF 5600 plus SCIEX.

Mass spectrometry is a technique which measures mass-to-charge ratios of ions. This can be used to identify and quantify the molecules of interest in the given sample. Since the advent of soft ionization methods, large biomolecules like proteins are increasingly being analyzed by mass spectrometry for studying various aspects of their structure. Mass spectrometry based Proteomics, i.e., large scale identification and quantification of proteins from a complex mixture of biological origin; and variations thereof, has become an important workflow in biological research. High resolution mass spectrometry is primarily applied for large scale discovery studies while low resolution mass spectrometry is more suited for targeted proteomics. At ACTREC, we have three high resolution mass spectrometers (MS) and one low resolution MS dedicated for omics applications.

Instruments
Cell
Cell

Orbitrap Exploris 480, Thermo

This high resolution MS system has an ESI ion source and a nanoFlex ion source and the mass analyzer is an Orbitrap. This system is coupled with a Vanquish nanoLC. The high resolution and high sensitivity provided by this system is useful in shotgun proteomics applications of discovery nature. There are specialized data analysis software suits for analysis of the raw data, namely Proteome Discoverer and Spectranaut. This system is well known for in-depth proteomic profiling of the sample, label and label-free quantification of the proteins in a complex mixture. This is well suited for comparative proteomics studies leading to biomarker identification for various disease conditions etc.

Cell
Cell

Synapt XS, Waters

This is a high resolution MS with an interesting option of changing the ion sources from ESI, nanoESI and MALDI (matrix assisted laser desorption ionization). For ESI and nanoESI application, it can be coupled with the M-Class UPLC from Waters and can be used for various shotgun proteomics, lipidomics, metabolomics applications. The main mass analyzer is TOF and the system is equipped with Ion Mobility Separation of the ions, providing separation between molecules of same molecular weight but different conformations. Since this system also has an option of Electron Transfer Dissociation (ETD), this is suitable for identification and characterization of the post-translational modifications (PTMs). With MALDI as an ion source, this system can be utilized for tissue MALDI imaging. The data analysis software available is Progenesis QIP and MassLynx.

Cell
Cell

Xevo TQ Absolute, Waters

This is a low resolution MS system with three quadrupole mass analyzers in series. It is coupled with an H-Class UPLC and has an ESI ion source. This system is best suited for targeted proteomics studies, where absolute or relative quantification of the protein/peptide/small molecule of interest can be done using single or multiple reactions monitoring technique (SRM/ MRM) and its variations. The unparalleled sensitivity provided by the MRM assay can be used to track extremely low amounts of compounds of interest in the given sample. The analysis software MassLynx software helps in analyzing the data of targeted quantification experiments

Cell
Cell

TripleTOF 5600 plus, SCIEX

This is a high resolution MS with nanoESI (electrospray ionization) source and nanoLC coupled with it. The main mass analyzer is Time-of-Flight (TOF). This system is useful for shotgun proteomic experiments of qualitative and quantitative nature. The patented SWATH technology of Sciex helps in increasing the depth of the proteomic identification as well as label-free quantification, and thereby able to identify the proteins with very low expression. Along with that, the system can also be used for label-based quantification of proteins. The analysis software ProteinPilot is useful for qualitative and quantitative proteomics studies.

Applications
Sr. No.Applications
1Enriched Proteins (IP / In-gel digested samples)
2Global Protein Profiling (Complex Mixture)
3Label Free Quantification
4Labelled Quantification (i-TRAQ / TMT)
5Parallel Reaction Monitoring (PRM)
6PTMs
7Metabolomics
8Lipidomics
9MALDI MS
10MALDI MS-MS
11Targeted Proteomics Studies
12SRM / MRM studies
Publications

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Staff
Sr. No.NameDesignationEmail ID/Phone/Extension
1Mr. Shashadhar DolasScientific Officer ‘E’ OIC-Mass Spectrometry FacilitySdolas[at]actrec[dot]gov[dot]in
5475/5320
2Dr. Heramb KulkarniScientific Officer ‘D’Heramb.kulkarni[at]actrec[dot]gov[dot]in
5475/5320/5394
3Mrs. Poonam KawleScientific Assistant ‘F’mass.spec[at]actrec[dot]gov[dot]in
5475/5320
4Mr. Prince Ragul V.Technician ’A’mass.spec[at]actrec[dot]gov[dot]in
5475/5320
Contact Details

Mass Spectrometry

KS-08 & KS-19, Khanolkar Shodhika,
Advanced Centre for Treatment, Research & Education in Cancer (ACTREC),
Tata Memorial Centre,
Sector-22, Kharghar,
Navi Mumbai – 410210,
Maharashtra, India.

Email : mass.spec[at]actrec[dot]gov[dot]in
Phone No: +91 (022) 27405000 / 68735000
Extension 5475/5320

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