| Course Name | Analytical Biochemistry |
| Course Code | 25BIO205 |
| Program | B. Sc. (Hons.) Biotechnology and Integrated Systems Biology |
| Semester | 3 |
| Credits | 3 |
| Campus | Amritapuri |
Protein extraction and quantitation: Enzymatic lysis, Homogenizer, Blender, Sonication, Bead mill shaker, French press, Biuret, Lowry, BCA and Bradford Assays. Protein precipitation and treatment: Salting-in, Salting-out, Effect of organic solvents and polymers, Dialysis, Ultrafiltration, Centrifugation.
Chromatography: Partition coefficient, Retention, Resolution, Gel filtration chromatography, Ion exchange chromatography, Affinity chromatography, Hydrophobic interaction chromatography, Hydroxyapatite chromatography, Paper chromatography, thin layer chromatography, Reversed-phase chromatography, Normal phase chromatography.
HPLC: Fundamentals of high-performance liquid chromatography, Columns, Detectors.
Electrophoresis: Native PAGE, SDS-PAGE, Isoelectric focusing, 2D-PAGE.
Spectroscopy: Fundamentals of UV/Vis Spectroscopy, Applications of UV/Vis spectroscopy, Spectrophotometer, Fundamentals of fluorescence spectroscopy, Jablonski diagram, Spectro fluorometer, Applications of spectrofluorimetric.
LEARNING OBJECTIVES:
The main objective of this course is to provide basic knowledge to students to understand analytical tools and apply them to decipher structure and functions of biomolecules.
COURSE OUTCOMES:
After completing the course, students shall be able to
CO1. Describe important biomolecular extraction, quantitation, separation and purification techniques.
CO2. Recall concepts and applications of UV/Visible and fluorescence spectroscopy.
CO3. Differentiate important techniques to analyze biomolecules.
CO4. Solve qualitative and quantitative problems related to biomolecular characterization.
REFERENCES:1. Protein Purification Techniques: A Practical Approach, Simon Roe, Oxford, 2nd Ed., 2004.2. Protein Purification: Principle and Practice, Robert K. Scopes, Springer, 3rd Ed., 1994.3. Physical Biochemistry: Principles and Applications, David Sheehan, John Wiley & Sons, 2nd Ed., 2000.4. Practical Biochemistry: Principles and Techniques, Keith Wilson and John Walker, Cambridge, 5th Ed., 2004.
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