NIT Jalandhar V Sem BIOTECHNOLOGY Syllabus
DEPARTMENT OF BIOTECHNOLOGY
Detailed syllabus 5th Semester:
BT 301 Separation Methods in Biotechnology [3 1 0 4]
Separation of insoluble products: sedimentation, sedimentation coefficient, filtration,
membrane filtration, centrifugation, microcentrifuge, ultracentrifuge, differential and density
gradient centrifugation, coagulation and flocculation.
Cell Disruption: Mechanical methods, Non-mechanical methods.
Dialysis and Filtration: electro-dialysis, ultra-filtration and micro-filtration, cross-flow ultrafiltration
Separation of soluble products: Liquid-liquid extraction, aqueous two-phase extraction,
precipitation, adsorption, salt and solvent precipitation of protein, recombinant protein
Electrophoresis: Gel electrophoresis (Agarose, PAGE, SDS PAGE), Disc gel electrophoresis,
Gradient electrophoresis, pulse field gel electrophoresis, 2 D gel electrophoresis, capillary
electrophoresis, Isoelectric focusing, Gel capillary electrophoresis, Capillary zone
electrophoresis, Autoradiography, Radioimmunoassay.
Chiral separation of biomolecules: Chiral Thin layer chromatography, chiral gas-liquid
chromatography, non chromatographic chiral separation
Chromatography: Method selection; selection of matrix; Adsorption chromatography, Ionexchange
chromatography, gel-filtration chromatography, size exclusion chromatography, ion
exclusion chromatography, affinity chromatography, hydrophobic interaction chromatography,
high pressure liquid chromatography, Co-valent chromatography; IMAC chromatography, Dye
ligand chromatography. Chromatography scale-up.
Crystallization: Theory and methods; API-electrospray and MADI-TOF; Mass spectrometry;
Enzyme and cell immobilization techniques; DNA & Peptide Synthesis.
Reverse Micelles: Reverse micelles formation, correlation of miceller size and protein size,
Reverse micelles extraction method
Molecular Imprinting: Imprint property, selectivity of molecular imprinting.
Drying: Lyophilization, Spray drying, vacuum drying, air drying.
1. Willard, H., Meritt,L.L., Dean J.A. and Settle F.A., “Instrumental Methods of analysis”,
6th edition, CBS Publishers, (1986)
2. Vogel’s, “Textbook of Quantitative Chemical Analysis”, 6th Edition, Pearson, (2005).
3. Skoog,D.A. , F.J. Holler and T.A. Nieman., “Principles of Instrumental analysis”, 5th
Edition , Harcourt Area PTE ( 1998)
4. Okotore, R.O., “Basic Separation Techniques in Biochemistry”, New Age (1998)
5. Sivasankar, B., “Bioseparation: Principles and Techniques”, Prentice Hall India (2005).
6. R. Scopes, Protein Purification – Principles & Practices, 3rd Edition, Springer Verlag,
BT-303 Animal Cell and Tissue Culture [3 0 0 3]
Basics of Cell and Tissue Culture: Laboratory requirements for tissue culture, substrates for
cultures, culture media for animal cell cultures, culture procedures and principles, freeze storing
of cells and transport of cultures, Primary culture, secondary culture; Continuous cell lines;
Characteristics of Cells in Culture: Contact inhibition, anchorage independence/dependence,
cell-cell communication, cell senescence.
Cell Culture Lines: Definition, development and maintenance, characteristics of animal cells
and their implication on process design, nutritional requirements and serum free culture of
mammalian cells, kinetics of growth and product formation, cloning of cell lines, cell
synchronization, viral sensitivity of cell lines, cell line characterization, stem cell lines.
General Tissue Culture Techniques: Types of tissue cultures, methods of disaggregating
primary cultures, primary tissue explanation technique, reactor systems for large-scale
production using animal cells.
Organ Culture: Methods, behavior of organ explants and utility of organ culture, whole embryo
Methods in Cell Culture: Micro carrier cultures, cell immobilization, animal cell bioreactor,
large scale cell cultures for biotechnology, somatic cell fusion, flow cytometry, transfection.
Applications of Animal Cell Culture: Use in gene therapy, cloning from short-term cultured
cells, cloning from long-term cultured cells, cloning for production of transgenic animals,
cloning for conservation. Application of animal cell culture for in vitro testing of drugs; Testing
of toxicity of environmental pollutants in cell culture.
Hybridoma technology: Production of monoclonal and polyclonal antibodies with different
types of antigens, antigen preparation and modification, adjuvants dose and route of antigen
administration, collection of sera, purification of antibodies, production and applications of
monoclonal antibodies for diagnosis and therapy, production of virus vaccines, specific vaccines,
production of cellular chemicals like Interferons, Interleukin etc. Immunoassay procedures.
1. Freshney R. Ian, “Culture of animal cells: A manual of Basic Technique”, Willey-Liss
Publisher, 5th edition (2005).
2. Jenkins N, ed., “Animal Cell Biotechnology: Methods and Protocol”, Humana Press (1999).
3. Minuth W.W., Strehl R., Schumacher K., “Tissue Engineering: Essential for Daily
Laboratory Works”, Willey Publisher (2005).
4. Butler, M “Mammalian Cell Biotechnology- A Practical Approach,” IRL Oxford University
BT-305 Separation Process Laboratory [0 0 2 1]
1. Harvesting of fermentation broth and its processing for product purification.
2. Solid-liquid separation
3. Liquid-liquid separation
4. Disruption of microbial cells
5. Separation by precipation through adding salts and solvents.
8. Vaccum evaporation
9. Drying and crystallization
10. Separation of proteins and other biomolecules by various Chromatography techniques
BT-307 Cell and Tissue Culture Laboratory [0 0 4 2]
1. Introduction to Tissue Culture Laboratory facilities
2. Preparation of medium and sterility tests
3. Principles and Technique for monolayer and suspension culture
4. Subculture of animal cell line and cell preservation
5. Genetically engineered cell
6. Mass cell cultivation
7. Preparation of Culture Media for plant cell, Sterilization of Culture Media
8. Explant selection, sterilization and inoculation;
9. Various media preparations; MS, B5, SH PC L-2;
10. Callus and cell suspension culture
11. Plant regeneration from embryo, meristem and callus culture.
BT-321 Plant Cell and Tissue Culture [3 1 0 4]
Introduction: Special features of plant cells, totipotency, regeneration of plants, organogenesis,
Somatic Embryogenesis, somaclonal variation, its genetic basis and application in crop
Basic techniques in cell and tissue culture: Culture media composition and preparation, cell
Cell Cultures: Initiation and maintenance of callus and suspension culture, protoplast isolation,
fusion and culture, somatic hybridization, Role of tissue culture in rapid clonal propagation,
production of pathogen – free plants and “synthetic seeds”. Overcoming Barriers using Tissue
Culture: Pre- and Post-Fertilization barriers, Production and Use of Haploids.
Micropropagation : Techniques, factors affecting morphogenesis and proliferation rate ,
technical problems in micro propagation, meristem culture for the production of pathogen free
plants , applications of micro propagation.
Protoplast technology: Isolation, culture and plant regeneration, protoplast fusion, identification
and characterization of somatic hybrids, applications of protoplast technology.
Biochemistry of major metabolic pathways and products: Autotrophic and heterotrophic
growth – carbon dioxide assimilation, carbohydrate metabolism, nitrogen assimilation.
Specific gene transfer: Indirect and direct methods, current status and limitations.
Plant products of industrial importance: Cell suspension culture development and production
of secondary metabolites by suspension cultures (case studies of azardiractin, podophyllotoxin)
Biological and technology barriers: Mutation, somaclonal variation, hydrodynamic shear and
its quantification, mixing and impeller design aspects.
Transgenic Plants: Genetically Modified Crops, Resistance against Biotic and Abiotic Stresses,
Plant Cell Reactors: Comparison of reactor performance, immobilized plant cell and cell
Automation in plant tissue culture: Field techniques for propagation of regenerated plants.
1. Bhojwani S.S. and Razdan M.K., “Plant Tissue culture Theory and Practice”,
Elesevier Science , Netherlands (2004)
2. Razdan M.K., “Introduction to Plant Tissue culture”, 2nd Edition, Science Publishers
3. Narayanswamy S.,”Plant Cell and Tissue culture”, Tata Mc-Graw Hill publishing Co.
4. Trigiano R.N., Grey D.J., “Plant Tissue Culture: Concepts and Laboratory Exercises”,
2nd Edition, CRC Press (2000).
5. Trigiano R.N., Grey D.J., “Plant development and Biotechnology”, CRC Press (2005).
6. Dixon R.A., Gonzales R.A., “Plant Cell Culture: A practical approach”, Oxford
University Press (1994).
BT-323 Agricultural Biotechnology [3 1 0 4]
Production of disease free plants : shoot – tip – cultures, shoot – tip – grafting, viricidal
Tissue culture as a source of genetic variability: somaclonal and gametoclonal variant
selection, sources and causes of variation, application in crop improvement.
Protoplast isolation: culture and fusion, selection of hybrid cells and regeneration of hybrid
plants, somatic hybridization
Plant cell cultures for useful chemicals: pigments perfumes, flavors, insecticides, anticancer
agents and pharmacologically important compounds.
Genetic Engineering in Agriculture: techniques for the insertion of foreign genes into plant
cells, Ti plasmid and vectors, production of transgenic plants, (i) Transgenic plants (ii) gene
cloning, restriction fragment length polymorphisms, transposons, and insertional mutagenesis.
Molecular Farming: Plants As factories for biopharmaceuticals, Transgenic value added
specialty crops, Use of antisense RNA and other technologies.
Nitrogen fixation: nif-gene transfer, herbicide resistance and stress tolerance in plants. Isolation
and characterization of organelle genome (Plastome and Chonodriosome).
Bioinsecticides and biofertilizers: Preservation of rare plant species germplasm collection and
conservation. Soil Reclamation: Phytoremediation
1. Bhojwani S.S. and Razdan M.K., “Plant Tissue culture Theory and Practice”, Elesevier
Science , Netherlands (2004)
2. Trigiano R.N., Grey D.J., “Plant Tissue Culture: Concepts and Laboratory Exercises”, 2nd
Edition, CRC Press (2000).
3. Lindsey K, “Plant Tissue culture Manual”, Kluwer Academic Publ. (1991).
4. Kung S D, Wu R, “Transgenic Plants Vol. 1 & 2”, Academic Press, San Diego (1993).
5. Lindsey K, Jones M G K, “Plant biotechnology In Agriculture”, Prentice hall (1990).
BT-325 Biochemical Reaction Engineering [3 1 0 4]
Biochemical Reaction Engineering: Kinetics of homogeneous reactions; reaction mechanism;
Temperature dependency from Arrhenius law; Theoretical prediction of rate constant:
Interpretation of batch kinetic data.
Kinetics of enzyme catalyzed reactions in free and immobilized states: Michaelis-Menten
equation and its various modifications. Effects of External mass transfer in immobilized enzyme
systems; analysis of intraparticle diffusion and reaction.
Kinetics of substrate utilization, product formation and biomass production : Monod
growth model and its various modifications; structured and unstructured kinetic rate models;
Thermal death kinetics of cells & spores; Transport phenomena in Bioprocess systems; gasliquid
mass transfer in cellular systems. Mass transfer for bubbles swarms;
Types of reactors: batch, plug flow reactor (PFR), continuous stirred rank reactors (CSTR),
fluidized bed reactor, bubble column, air life Fermentor, etc; Concept of ideal and non-ideal
reactor: residence time distribution; Operating considerations in bioreactors for suspension and
immobilized cultures, modifying batch and continuous reactors, immobilized cell systems, solid
Models of non-ideal reactors: plug flow with axial dispersion, tanks-n-series model.
Unconventional bioreactors: Hollow fiber reactor, membrane reactor, perfusion reactor for
animal and plant cell culture.
Kinetics of mixed cultures: Major classes of interaction in mixed cultures, models describing
mixed-culture interactions, reaction dynamics, industrial application of mixed cultures.
1. Levenspiel O, “Chemical Reaction Engineering”, 3rd Ed , John Wiley & Sons, Singapore
2. Lee J M, “Biochemical Engineering” , Prentice Hall (1992)
3. Shuler M L, Kargi F, “ Bioprocess Engineering- Basic Concepts” , 2nd ed, Prentice Hall
of India Ltd. ( 2002)
4. Aiba S, Humphrey A E and Millis N F ,“Biochemical Engineering” , Academic Press
BT-327 Biosensor [3 1 0 4]
Overview of Biosensors: Fundamental elements of biosensor devices, Fundamental engineering
aspects of biosensors, Signal processing for biosensors.
Fundamentals of measurement science: applied to optical, electrochemical, mass, and pressure
Theoretical analysis of biosensor: design and performance.
Electro chemical biosensors: Electrochemical principles, Amperometric biosensors and charge
transfer pathways in enzymes, Glucose biosensors, engineering electrochemical biosensors,
Other than electrochemical or optical sensing schemes.
Optical Biosensors: Optics for biosensors, Attenuated total reflection systems, Non-invasive
Mass and Acoustic Biosensors: Saubrey formulation, Acoustic sensor formats, Quartz crystal
microbalance, Whole cell biosensors
1. Anthony E G C, Cooper J M, “Biosensors”, Oxford University Press (2004)
2. Roger K R , Mulchandani A, “Enzyme and Microbial biosensors”, Humana Press
3. Bilitewsk U, Turner A P F, “ Biosensor in Environmental Monitoring”, Taylor &
4. Donald G B, “Biosensors: Theory and Applications”, CRC Press (1993)
5. Donald L W, “Bioinstrumentation and Biosensors”, CRC Press (1991)
6. Donald L W, Wingard L B, “Biosensors with fiber optics”, Humana Press (1991