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NPTEL

Introduction to Proteomics

NPTEL via Swayam

Overview

This course introduces to the basic biology of proteins and the new advanced science called as proteomics which aims to look into the protein properties from a global perspective, i.e., not undertaking one protein at a time, but an entire set of proteins in the milieu. The course will cover in detail the two major aspects of proteomics i.e., Gel-based proteomics and Mass spectrometry-based proteomics. The gel-based module will cover different techniques like SDS-PAGE, 2-DE, 2D-DIGE etc. These techniques had a major contribution in transition from protein chemistry to proteomics. Mass spectrometry, on the other hand, is an advanced analytical technique for accurate mass measurement. In this module, we will discuss the basics of mass spectrometry, sample preparations, liquid chromatography, hybrid mass spectrometers and quantitative proteomics techniques such as iTRAQ, SILAC and TMT using mass spectrometry. The course will also provide the basic knowledge about sample preparation, mass spectrometry workflow, different chromatography technologies and quantitative proteomics.INTENDED AUDIENCE :It would be applied to B.Sc., M.Sc. and MS.PREREQUISITES : Any B.Sc. Or M.Sc.The target audiences of this course are required to have a basic introduction to biology.

Syllabus

Week 1 : Basics of Proteins and Proteomics

Lecture 1 : Introduction to amino acids
Lecture 2 : Introduction to Proteins
Lecture 3 : Protein folding & misfolding
Lecture 4 : Introduction to Proteomics
Lecture 5 : Lab session – Protein-protein interaction using label-free biosensors
Week 2 : Gel-based proteomics

Lecture 6: Sample preparation and pre-analytical factors
Lecture 7 : Sample preparation: Pre-analytical factors (contd.)
Lecture 8 : Sample preparation: Protein extraction and quantification
Lecture 9 : One-dimensional electrophoresis
Lecture 10 : Introduction to 2-DE

Week 3 : Two-dimensional gel electrophoresis (2-DE)

Lecture 11 : 2-DE: Second dimension, staining & destaining
Lecture 12 : 2-DE: Gel analysis
Lecture 13 : 2-DE Applications
Lecture 14 : 2-DE Applications (contd.) & Challenges
Lecture 15 : Lab session - Protein/peptide pre-fractionation using OFFGEL FRACTIONATOR & data analysis

Week 4 : Difference in gel electrophoresis (DIGE) & Systems Biology

Lecture 16 : 2D-DIGE: Basics
Lecture 17 : 2D-DIGE: Data analysis
Lecture 18 : 2D-DIGE: Applications
Lecture 19 : Systems biology and proteomics – I
Lecture 20 : Systems biology and proteomics - II

Week 5 : Basics of mass spectrometry

Lecture 21 : Fundamentals of mass spectrometry
Lecture 22 : Chromatography technologies
Lecture 23 : Liquid chromatography
Lecture 24 : Mass spectrometry: Ionization sources
Lecture 25 : Mass spectrometry: Mass analyzers

Week 6 : Basics of mass spectrometry and sample preparation

Lecture 26 : MALDI sample preparation and analysis
Lecture 27 : Hybrid mass spectrometry configurations
Lecture 28 : Lab session - Demonstration of Q-TOF MS technology
Lecture 29 : In-gel & in-solution digestion
Lecture 30 : Lab session - Sample preparation: tissue sample preservation technology

Week 7 : Quantitative proteomics

Lecture 31 : Introduction to quantitative proteomics
Lecture 32 : SILAC: In vivo labelling
Lecture 33 : iTRAQ: In vitro labelling
Lecture 34 : TMT: In vitro labelling
Lecture 35 : Quantitative proteomics data analysis

Week 8 : Advancement in Proteomics

Lecture 36 : Proteomics applications
Lecture 37 : Challenges in proteomics
Lecture 38 : OMICS and translational research
Lecture 39 : Lab session – Targeted proteomics using triple quadrupole mass spectrometry
Lecture 40 : Lab session – Targeted proteomics: multiple reaction monitoring

Taught by

Sanjeeva Srivastava

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