Page 12 - REC :: M.Tech. BioTech Curriculum and Syllabus - R2019

BY19111 PREPARATIVE AND ANALYTICAL TECHNIQUES IN Category L T P C
BIOTECHNOLOGY
0 0 4 2

Objectives:
 To prepare the student in all the latest preparative and analytical techniques required in research or Industry.

List of Experiments
1 Preparation of Acetate, Tris and Phosphate Buffer. Validation of Henderson Hasselbachequation.
Reactions of amino acids – Ninhydrin, Pthalaldehyde, Dansyl chloride – measurement using colorimetric and
2
fluorimetric methods.
Differential estimations of carbohydrates – reducing vs non-reducing, polymeric vs 13 oligomeric, hexose vs
3
pentose.
4 Estimation of protein concentration using Lowrys’ method and Dye-bindingmethod.
5 DNA determination by UV-Vis Spectrophotometer – hyperchromic effect.
6 Separation of lipids by TLC.
7 Enzyme Kinetics: Direct and indirect assays – determination of Km, Vmax and Kcat, Kcat/Km
8 Enzyme Immobilisation studies.
9 Ion-exchange Chromatagraphy – Purification of IgG andAlbumin
10 Gel filtration – Size based separation of protein.
11 Affinity chromatography – IMAC purification of His-tagged recombinantprotein
12 Assessing purity by SDS-PAGE GelElectrophoresis
Total Contact Hours : 90

Course Outcomes:
Having learned all the techniques in this lab, the student will become capable in enzymology, techniques
 required in the quantitation of biomolecules, downstream processing and the chemical modification of
proteins, which will prepare him for a career in research or employment in the biotech Industry.

Web links for virtual lab (if any)
Biochemical Methods: A Concise Guide for Students and Researchers, Alfred Pingoud, Claus Urbanke, Jim
1
Hoggett, Albert Jeltsch, 2002 John Wiley & Sons Publishers,Inc,
Biochemical Calculations: How to Solve Mathematical Problems in General Biochemistry, 2nd Edition, Irwin
2
H. Segel, 1976 John Wiley & Sons Publishers,Inc,
Principles and Techniques of Practical Biochemistry- Wilson, K. and Walker, J. CambridgePress
3

BY19201 BIOSEPARATION TECHNOLOGY Category L T P C
3 0 0 3

Objectives:
Understand the methods to obtain pure proteins, enzymes and in general about product development

R &D
 Have depth knowledge and hands on experience with on Downstream processes

UNIT-I DOWNSTREAM PROCESSING 8
Introduction to downstream processing principles characteristics of biomolecules and bioprocesses. Cell
disruption for product release – mechanical, enzymatic and chemical methods. Separation characteristics of
proteins and enzymes – size, stability, properties – Flocculation and conditioning of broth – Process design
criteria for various classes of bio products (high volume, low value products and low volume, high value