BY4211 Preparative and Analytical Techniques in Biotechnology Syllabus:
BY4211 Preparative and Analytical Techniques in Biotechnology Syllabus – Anna University PG Syllabus Regulation 2021
COURSE OBJECTIVES
1 To learn and understand the principles behind the qualitative and quantitative estimation of biomolecules
2 To gain hands-on experience in spectroscopic methods
3 To acquire practical experience by performing chromatography for the recovery and subsequent purification of target biological products.
LIST OF EXPERIMENTS
1. Preparation of Acetate, Tris and Phosphate Buffer. Validation of Henderson Hasselbach equation.
2. Reactions of amino acids – Ninhydrin, Pthalaldehyde, Dansyl chloride – measurement using colorimetric and fluorimetric methods.
3. Differential estimations of carbohydrates – reducing vs non-reducing, polymeric vs 13 oligomeric, hexose vs pentose.
4. Estimation of protein concentration using Lowrys’ method, Dye-binding method.
5. DNA determination by UV-Vis Spectrophotometer – hyperchromic effect. Separation of lipids by TLC.
6. Enzyme Kinetics: Direct and indirect assays – determination of Km, Vmax and Kcat, Kcat/ Km.
7. Restriction enzyme – Enrichment and unit calculation.
8. Ion-exchange Chromatagraphy – Purification of IgG and Albumin
9. Gel filtration – Size based separation of proteins
10. Affinity chromatography – IMAC purification of His-tagged recombinant protein
11. .Assessing purity by SDS-PAGE Gel Electrophoresis
12. Chemical modification of proteins – PITC modification of IgG and Protein Immobilization
TOTAL: 90 PERIODS
COURSE OUTCOMES
After completion of the course the students will be able to
CO1 To learn about the principles of buffer preparation, the reactions of amino acids and DNA determination
CO2 To understand the principles of quantitative estimation of carbohydrates using the spectroscopic method
CO3 To gain knowledge about principles and practical experience in the estimation of protein concentration
CO4 To know the principles of determining the kinetic parameters of an enzymatic reaction using direct and indirect assays
CO5 To acquire hands-on experience on purification processes through chromatography and to design processes for the recovery of target biological products
REFERENCES:
1. Pingoud, A., Urbanke, C., Hoggett, J. and Jeltsch, A., “Biochemical Methods: A Concise Guide for Students and Researchers”, Wiley-VCH, 2002.
2. Segel, I.H., “Biochemical Calculations: How to Solve Mathematical Problems in General Biochemistry”, 2nd Edition, John Wiley & Sons, 2004.
3. Wilson, K. and Walker, J., “Principles and Techniques of Biochemistry and Molecular Biology”, 7th Edition, Cambridge University Press, 2010.