BM 501 Essentials of Biophysics

L T P Hr Cr Marks
3 0 2 5 4 200

Objectives of the course:

To understand the principles of biophysics, biochemical techniques and instrumentation

Learning Outcome:

Apply the knowledge of instrumentation for medicine

Prerequisite:

Physics and basic knowledge of biology

Course Description:

Module 1 Contents (2 hours)

Blotting Techniques: Types of blotting techniques- Northern, Southern, Western,
Immunoblotting. Principle of blotting and detection, method and application. Advantages and
disadvantages.

Module 2 Contents (7 hours)

Chromatography: Concept of Chromatography, Adsorption Chromatography, Ion Exchange
Chromatography, Gel Chromatography, HPLC, Affinity Chromatography, Gas
Chromatography.

Module 3 Contents (4 hours)

Electrophoresis: Moving boundary electrophoresis, zone electrophoresis, different supports
used for electrophoresis, electrophoresis under native and denaturing conditions, occurrence of
artefacts, activity staining, isoelectric focussing, 2D gel electrophoresis.

Module 4 Contents (4 hours)

Centrifugation: Basic Principle of Centrifugation, Instrumentation of Ultracentrifuge
(Preparative, Analytical), Factors affecting Sedimentation velocity, Standard Sedimentation
Coefficient, Centrifugation of associating systems, Rate-Zonal centrifugation, sedimentation
equilibrium Centrifugation.

Module 5 Contents (5 hours)

X-Ray Crystallography – X-ray diffraction, Bragg equation, Reciprocal lattice, Miller indices
& Unit cell, Concept of different crystal structure, determination of crystal structure [concept
of rotating crystal method, powder method].

Module 6 Contents (5 hours)

Spectroscopy: Raman Spectroscopy – What is Raman effect, Quantum mechanical reason of
Raman effect, Molecular Polarizability, Polarizability ellipsoid, Experimental technique of
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Raman effect, Basic concept of Pure Rotational & Vibrational, Raman spectra of simple
molecule (linear molecule). NMR Spectroscopy – Basic principle of NMR spectroscopy,
Experimental technique & instrumentation, Chemical shift, Hyperfine splitting, Relaxation
process. Absorption Spectroscopy – Simple theory of the absorption of light by molecules,
Beer-Lambert law, Instrumentation for measuring the absorbance of visible light, Factors
affecting the absorption properties of a Chromophore.

Module 7 (3 hours)
Radiation Biophysics: Radioactive labeling & counting, Autoradiography, Properties of
radioisotopes, commonly used isotopes, Use of isotopes in biology. G.M, counter, isotopic
dilution analysis, Scintillation Detectors. Radiation hazards. Effect of radiation on Nucleic
acids, Proteins, Enzymes, Cellular effects of radiation (somatic & genetic effects, Inhibition of
Mitosis, acute and chronic (whole body) effects of radiation).

Books/References:
1. Principles and Techniques of Biochemistry and Molecular Biology, 6th edition (2008),
2. Keith Wilson and John Walker, Publisher–Cambridge University Press.
3. Biophysics, Vasant Pattabhi, Gautam (2002), Narosa
4. Biophysics, Hopp, Lohman, Mark and Ziegler 3.4 Advances in Biophysics, Vol 18, 15
3.5 Molecular and Cellular Biophysics, Meyer B Jackson (2006), Cambridge)
5. Biomembrane structure and Function, Chapman D.
6. Biophysics by W. Hoppe. et. al., Springer - Verlag, 1989.
7. Essentials of Biophysics by P. Narayanan, New Age International (P) Ltd. Publishers,
New Delhi, 2000.
8. Radiation Biophysics by L.Alpen Edward, Academic Press, (1988).
9. Knoll, Glenn F. Radiation detection and measurement1Glenn F. Knoll. - 3rd ed.(1999)

BM501 Practical in Biophysics:

1. To study the effect of different solvents for a given pure compound using thin layer
chromatography.
2. To locate enzymes on electrophoreogram by active staining.
3. To separate protein by gel filtration and estimate the molecular weight.
4. To separate protein by ion exchange chromatography.
5. To separate protein by affinity chromatography.
6. Effect of UV rays on bacterial cell growth to evaluate LD50 and EC50.
7. To plot absorption spectrum of DNA and protein and find λmax using
spectrophotometer.
8. Visit to X-Ray Crystallography, NMR facility.

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