18BT55 Bioanalytical Techniques syllabus for BT

INTRODUCTION:

Extraction, Pre-treatment, Stabilization and preparation methods of bioproducts for analysis. Electrophoretic Techniques: Principle, equipment and process, Agarose gel electrophoresis, gradient electrophoresis, horizontal and vertical gel electrophoresis, electrophoresis techniques, isoelectric focusing, capillary electrophoresis and application of electrophoresis in analyzing macromolecules. Chromatography – principles, instruments and practice, adsorption, reverse phase, ion exchange, size exclusion, hydrophobic interaction, bio-affinity and pseudo affinity chromatographic techniques

CHROMATOGRAPHY AND PURIFICATION:

Chromatography: Classification of chromatographic techniques and their principles, Theory of chromatography, band broadening, rate and plate theory factors responsible for separation. Column chromatography, TLC, Paper chromatography.

Liquid Chromatography and HPLC: Instrumentation, pumps, solvent delivery system, isocratic and gradient programming modes, sample introduction system, columns, detectors, reversed phase and normal phase chromatography.

Gas Chromatography: Instrumentation, carrier gas supply, injectors, columns, packed and capillary columns, column oven and temperature programming, different detectors. Introduction to hyphenated techniques in chromatography, GC-MS and LC-MS.Cell fractionation and flow cytometry

SPECTROSCOPY:

UV-Visible spectroscopy, Fluorescence spectroscopy, IR/Raman spectroscopy,NMR Spectroscopy and Mass spectroscopy Types of nuclear magnetic resonance Chemical shifts – spin – spin coupling – relaxation mechanisms – nuclear overhauser effect –ESR multidimensional nmr spectroscopy – determination of macromolecular structure by NMR –magnetic resonance imaging.

SPECTROMETRY:

Ion sources sample introduction – mass analyzers and ion detectors – bimolecular mass spectrometry – peptide and protein analysis – carbohydrates and small molecules – specific applications. X-Ray diffraction: Scattering by x- rays – diffraction by a crystal – measuring diffraction pattern – Bragg reflection –unit cell – phase problem – anomalous diffraction – determination of crystal structure – electron and neutron diffraction.

INSTRUMENTAL METHODS OF ANALYSIS:

Advanced Microscopy: Morphology and identification of cells using microscopic studies like SEM, TEM and Confocal Microscopy.

EDAX, Elemental mapping, FTIR, UV-Visible spectrophotometer, Nanomechanical Characterization using Nanoindentation, Differential Scanning Calorimeter (DSC), Differential Thermal Analyzer (DTA), Thermo gravimetric Analysis (TGA), TEM, X-ray Photoelectron Spectroscopy (XPS), Electrochemcial Characterization measurements.

Course outcomes:

At the end of the course the student will be able to:

• Define the fundamentals of downstream processing for product recovery
• Understand the requirements for successful operations of analytical techniques
• Apply principles of various analytical devices used in in research and enhance problem solving techniques

Question paper pattern:

• The question paper will have ten full questions carrying equal marks.
• Each full question will be for 20 marks.
• There will be two full questions (with a maximum of four sub- questions) from each module.
• Each full question will have sub- question covering all the topics under a module.
• The students will have to answer five full questions, selecting one full question from each module.

Textbook/s

1 Analytical techniques in biochemistry and molecular biology R. Katoch Springer, New York, 2011

2 Biological spectroscopy I. D. Campbell Benjamin/Cummings Pub. Co 1984

3 Separation Processes in Biotechnology Asenjo, Juan A. CRC / Taylor & Francis 1990

Reference Books

1 Modern experimental biochemistry R. F. Boyer Benjamin Cummings 3rd ed., 2000

2 Principles of fluorescence spectroscopy J. R. Lakowicz Springer, New York 2006