Syllabus

The course contains a series of 8-10 experiments employing different analytical tools to characterize purified molecules. The design of the experiments is to give the students first-hand experience with expression/extraction and characterization of biomolecules/small bioactive molecules of interest.
Module I: Expression of recombinant human protein (CPL-1/Synuclein), and the analysis of the expression using UV-visible spectroscopy (for IPTG induction), PAGE (for expression of different proteins in the host).

Purification of the expressed protein using Ion Exchange Chromatography.

Structural dynamics in purified protein (time/additive-based) using steady-state fluorescence spectroscope (including extrinsic/intrinsic fluorescence and fluorescence anisotropy).

Structural dynamics in purified protein (time/additive-based) using Circular Dichroism Spectropolarimeter.

Thermodynamics of protein binding to different moieties/surfaces using Isothermal Titration/Differential Scanning Calorimeter(s).

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

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

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:
Categories different studied techniques as per their underlying principle and utilize them for designing the experiment/project,

Tailor different specifications of the instrument as per the needs of the project,

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 , x

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

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