Syllabus

The course contains a series of 8-10 experiments employing different tools for biophysical characterization of a purified biomolecule. The design of the experiments is to give the students first-hand experience with the characterization of biomolecules/small bioactive molecules of interest.
Module I: a. Buffer preparation of pH 4, 6.5 and 7.

Module II: b. Direct method of the purified protein estimation,
c. Indirect method of the purified protein estimation,
d. Demonstration of Autoclave and Sonicator for processing of used culture for recombinant protein expression,
e. Dialysis and Lyophilization of purified protein for further experiments.
f. Comparison of protein concentrations obtained after dialyzing the sample versus a non-dialysed sample (post-lyophilization)

Module III: g. Collection of thermogram of the purified protein at different pH buffers using Differential Scanning Calorimeter.
h. Interpretation of different thermodynamics parameters from the thermogram.
i. Comparison of the thermograms of the protein collected in different conditions.

Module IV: j. Secondary structure estimation of the purified protein using circular dichroism spectropolarimeter.
k. Characterization of different bonds present in purified biomolecule sample using ATR-FTIR Spectroscope.

The main objective, in brief, of the course is to give the students hands-on experience with different biophysical tools/methods employed in pharmaceutical/biotechnological industries and academic groups working in biological sciences. Hence, the specific objectives of the course are to:
1. Familiarize the students with the handling of analytical instruments and data analysis,

2. Understand different thermodynamic parameters’ role in governing different structures (molecular/population heterogeneity) to biomolecules,

3. Impart knowledge necessary to design and interpret the experiment's outcome using studied analytical tools.

After completion of the course, the student will be able to:
a. Categories different studied techniques as per their underlying principle and utilize them for designing the experiment/project,

b. Tailor different specifications of the instrument as per the need of the project,

c. Understand the strengths and limitations of the techniques and use them in the design, execution, and interpretation of the outcomes from the experiment.

Keith Wilson, John Walker, Practical Biochemistry (Principles and techniques),, Cambridge University Press,

Arrondo JLR, Alonso A (Eds.), Advanced Techniques in Biophysics, Springer (Series in Biophysics)

J. A. Glasel, and M. P. Deutscher, Introduction to Biophysical Methods for Protein and Nucleic acid Research, Academic Press