Download M.Tech. Industrial BioTechnology Syllabus
BIOPHARMACEUTICALS
Subject Code : 14IBT153
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To understand fundamental principles of drug development methods and to describe various procedures involved in drug development and pharma operations. To learn operations and practices followed in biopharma industry. To learn about and describe the various pharma products of microbial and animal origin. To understand the mechanism and functioning of biopharmaceutical products. To apply methods of recombinant DNA technology for the production of biopharmaceuticals. To learn methods of quality assurance and validation procedures in biopharama sector.
Course Outcomes: At the end of this course, student will be able to:
· Demonstrate fundamental principles of drug development methods.
· Understand operations and practices followed in biopharma industry.
· Describe various procedures involved in drug development and pharma operations.
· Explain production of various pharma products of microbial and animal origin.
· Describe mechanism and functioning of biopharmaceutical products.
· Apply methods of recombinant DNA technology to production of biopharmaceuticals.
· Identify and demonstrate methods of quality assurance and validation procedures in biopharama sector.
MODULE 1 DRUG DEVELOPMENT 10 Hours
Medicinal plants as source for new drug development – Screening techniques for natural products – Lead optimization – Structure-activity studies through computer-aided modeling – parallel synthesis and combinatorial libraries – rapid screening of combinatorial libraries.
MODULE 2 PHARMACEUTICAL OPERATIONS & PRACTICE 10 Hours
Principles and equipment for: Extraction, drying, evaporation, distillation, centrifugation, filtration, comminution, particle sizing, powder handling, granulation, pelletization, coatings Pharmacopoieas, Formulations and Legislations – Pharmaceutical Calculations
MODULE 3 MICROBIAL & ANIMAL PRODUCTS 10 Hours
Vaccines and sera: Bacterial vaccines, Viral vaccines – antibiotics: penicillins, cephalosporins, chloramphenicol, tetracyclines, erythromycin, novobiocin, fusidic acid, vancomycin, rifampcin, polypeptide antibiotics, carbohydrate antibiotics – Bacteriocins – blood products and plasma substitutes: plasma, serum and plasma fractions, fractionation of plasma, coagulation factors, human immunoglobulin, albumin preparations, plasma substitutes – disinfection: disinfectants, mode of action, factors influencing disinfectant action – sterilization: sterilization processes, preparation of sterile medicaments
MODULE 4 PHARMACEUTICALS 10 Hours
Recombinant DNA products: Human insulin, interferon, somatostatin, somatotropin, streptokinase – recombinant bioconversions of bioactive molecules: production of 7-
aminocephalosporanic acid from cephalosporin, chemoenzymatic production of Epivir™ – Pharmacogenomics – Genetic polymorphisms in: drug metabolism, drug transport, drug targets
MODULE 5 VALIDATION TECHNIQUES 10 Hours
Validation Techniques for pharmaceutical industries Pilot Plant Scale-Up Techniques Analytical methods and tests for various drugs and Packaging techniques – Glass containers, plastic containers, film wrapper, bottle seals. Quality assurance and control. Quality control in clinical trials; Monitoring and audit; Inspections; Pharmacovigilance; Research governance; Trial closure and pitfalls-trial closure; Reporting and legal requirements; Common pitfalls in clinical trial management.
TEXT / REFERENCE BOOKS
1. E.A. Rawlins (Ed.), “Bentley’s Textbook of Pharmaceutics”, Bailliere Tindall, UK, 8th Ed., 1992.
2. Leon Shargel, Susanna Wu-Pong, Andrew B.C. Yu, “Applied Biopharmaceutics & Pharmacokinetics”, Fifth edition, McGraw Hill, 5th Ed., 2005.
3. Proceedings of the Workshop on “Developments in Drugs & Pharmaceutical Technology”, Organized by NAM S&T Centre, Ed: J.N. Mishra, Daya Publishing House, 2004.
4. A.R. Gennaro (Ed.) “Remington: The Science & Practice of Pharmacy”, Volumes I & II, Indian Edition, Lippincott Williams & Wilkins, 2004.
5. Gray Walsh & B. Murphy, “Biopharmaceuticals and industrial prospective”, Kluwer publishers 1999.
6. Gray Walsh, “Biopharmaceuticals”, Wiley John & Sons, Inc. 2003.
7. Camille G. Wermuth, “The practice of Medicinal chemistry”, Academic Press, 2003.
8. Dann, J.A, Crommelin & Robert D., Sindelar, “Pharmaceutical Biotechnology”, Taylor & Francis, 2002.
9. Nallari, P. and Rao, V. V. “Medical Biotechnology”, Oxford University Press, 2010.
FERMENTATION TECHNOLOGY – I
Subject Code : 14IBT12
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn various cell culture methods, strain improvement and to design and develop medium for inoculum development; To understand techniques of sterilization and to study the various aspects of fermenter for an industrial fermentation process; To apply the knowledge of control system for control of industrial fermentation process.
Course Outcomes: At the end of this course, student will be able to:
· Demonstrate the methods of cell culture under various conditions, strain improvement methods
· Design and develop medium for cell cultivation for fermentation process
· Apply the knowledge of sterilization techniques
· Understand needs of various parts of fermenter and their operation
· Apply the knowledge of control theory for industrial fermentation control
MODULE 1 CELL CULTIVATION AND GROWTH KINETICS 10 Hours
Cell culture (Bacteria, fungal, plant, animal), Microbial growth kinetics, logistic growth model, growth of filamentous organism Strain improvement of industrial micro organism.
Measurement of cell mass. Cell immobilization. Numericals.
MODULE 2 INOCULUM DEVELOPMENT AND MEDIA PREPARATION 10 Hours
Media components and optimization (PB, RSM techniques), types of media, Strain preservation , inoculum preparation, Development of inocula for industrial fermentation/ seed fermenter.
MODULE 3 STERILIZATION 10 Hours
Sterilization: death kinetics, del factor, batch and continuous; insitu and ex-situ sterilization, Sterilization of medium, air, filters, fermenter. Numericals.
MODULE 4 FERMENTATION PROCESS 10 Hours
Parts of fermenter: Body, Baffles, Sparger, valves, ports, Aeration: Oxygen requirement, Oxygen uptake in cell culture, Oxygen transfer in fermenter, gas hold up, KLa measurement, Measurement of dissolved oxygen concentrations, Estimating Oxygen Solubility, Measurement of KLa, factors effecting KLa in fermenter, Agitation: fluid rheology. Numericals.
MODULE 5 CONTROL OF INDUSTRIAL FERMENTATION 10 Hours
Requirements for control, sensors, controllers, design of fermenter control specification, control of incubation, advanced incubation control.
TEXT BOOKS:
1. Stanbury, Vitaker and Hall, “Principles of Fermentation Technology”, Butterworth Heinemann, 2nd Ed., 1999.
2. El-Mansi (Ed.), “Fermentation Microbiology and Biotechnology”, CRC Press, 3rd Ed.,2011.
REFERENCE BOOKS:
1. Pauline M. Doran, “Bioprocess Engineering Principles”, Academic Press, 2nd Ed., 2012.
2. Badal C. Saha (Ed.), “Fermentation Biotechnology”, CBS Publishers & Distributors Pvt. Ltd., 2004.
3. Brian McNeil and Linda Harvey (Ed), “Practical Fermentation Technology”, Wiley, 2008.
ADVANCED MOLECULAR BIOLOGY
Subject Code : 14IBT13
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn and understand procedures in molecular biology research to work with nucleic acid processing; To gain the knowledge of gene expression models of prokaryotic and eukaryotic system; To apply the knowledge of molecular research in rDNA technology and in therapeutics; To use fundamental experimental knowledge of molecular research procedures in understanding molecular biology concepts, detection and therapy.
Course Outcomes: At the end of this course, student will be able to:
· Demonstrate working procedures and protocols in molecular research
· Understand gene expression models
· Apply molecular research concepts in rDNA technology and in therapeutics
· Analyze and know the requirements of vectors and protein expression
· Design recombinant vectors for therapeutic applications
MODULE 1 MOLECULAR RESEARCH PROCEDURES AND WORKING WITH NUCLEIC ACIDS 10 Hours
Chemical synthesis of DNA[Glick], synthetic genes, isolation of DNA, RNA, handling and quantification of nucleic acids, labeling, nucleic acid hybridization. PCR: essential features, designing of primers, DNA polymerases of PCR, exotic PCR techniques (PCR using mRNA (RT-PCR), nested PCR, inverse PCR, RAPD, processing of PCR products, applications.
Alternative amplification techniques, Production of gene probes: gene probe labeling, non radioactive DNA labeling, end labeling of DNA, labeling by primer extension, nick
translation labeling. Nucleotide sequencing: Maxam Gilbert, Sanger method, direct PCR sequencing, cycle sequencing, automated flourosense DNA sequencing (primer walking).
MODULE 2 GENE EXPRESSION IN PROKARYOTES AND EUKARYOTES AND MANIPULATION OF GENE EXPRESSION 10 Hours
Prokaryotes; Prokaryotic gene expression and control of gene expression; isolation of functional promoters: Promoter selection with E. coli plasmid pBR316 and pK01. Gene expression from strong and regulatable promoters: Regulatable promoters, increasing protein production, large scale systems, expression in other microorganisms. Fusion proteins: cleavage of fusion proteins, uses of fusion proteins, expression of native protein, DNA integration into host chromosome.
Eukaryotes: some considerations in choice of cell lines, endogenous selectable markers and dominant selectable markers, stepwise amplification of transgene, plasmid vectors for transfection, major expression systems used in animal cells.
MODULE 3 rDNA TECHNOLOGY 10 Hours
Early thoughts and experiments in cloning, first step towards cloning frogs and toads, nuclear totipotency,
Prokaryotic vectors: Bacterial plasmids, viral vectors: cosmids, phasmids, M13 vectors, broad host range vectors.
Eukaryotic vectors: Generalized eukaryotic expression vector, Yeast expression systems: Saccharomyces cerevisiae vectors, yeast selectable markers, direct expression in
Saccharomyces cerevisiae, secretion of heterologous proteins by Saccharomyces cerevisia; Other yeast expression systems: Expression of hepatitis B virus surface antigen, expression of bovine lysozyme C2, cloning of large DNA fragments in BAC and YAC vectors; cultured insect cell expression system: Baculovirus transfer vector, Scaleup problem with Baculovirus system; Mammalian cell line expression system: Human Papova BK virus shuttle vector, Production of protein drug for clinical trials, viral vectors-adenovirus, retrovirus, pox virus and bacculovirus.
Plant as bioreactors: biopharming and neutraceuticals (edible vaccines, Ab, polymer producers from plants). Live recombinant vaccines.
MODULE 4 GENE EXPRESSION DIRECTED MUTAGENESIS AND PROTEIN ENGINEERING 10 Hours
Oligonucleotide directed mutagenesis with M13 DNA, PCR amplified oligonucleotide directed mutagenesis, degenerate oligonucleotide primers, random mutagenesis and site directed mutagenesis. Adding disulphide bonds, changing aspargine to other amino acids, reducing number of free sulphahydril residues, increasing enzyme activity, modifying enzyme specificity, increasing protein stability.
Applications: Point mutation- Interferons β16(betaseron/ betaferon), lispro insulin(humalog), novel vaccine adjutants, domain shuffling, linking domains, swapping protein domains, deleting domain , whole protein shuffling, fusion proteins.
MODULE 5 rDNA TECHNOLOGY FOR PRODUCTION OF THERAPEUTICS 10 Hours
Recombinant interferon. Subunit vaccines: against herpes simplex virus, foot and mouth disease, peptide vaccines. Live recombinant vaccines: Vaccinia virus recombinants, BCG vaccines, poliovirus chimaeras.
Attenuated vaccines: Cholera, Salmonella as live bacterial vaccine
Vector vaccines: vaccines directed against virus and bacteria
Monoclonal antibodies: Isolation of immunoglobulin variable region genes and expression on the surface of bacteriophage- isolation of mRNA for VH and VL and generation of cDNA, PCR amplification of cDNA for antibody VH and VL. Linking of VH and VL to give scFv, Insertion of scFv into phagemid vector, expression of scFv on the surface of bacteriophage, screening phage display libraries of immunoglobulin genes, preparation of soluble scFv, screening supernatants containing soluble scFv, application of monoclonal antibodies in biomedical research, diagnosis and treatment of diseases.
TEXT BOOKS:
1. Primrose, S. B and Twyman, R. M. “Principles of gene manipulation and genomics”. Blackwell Publishing, 7th Ed, 2006.
2. Gerald Karp. “Cell and Molecular Biology”. John Wiley, 6th Ed., 2009.
REFERENCE BOOKS:
1. Walker, J. M. and Rapley, R. “Molecular Biology and Biotechnology”. Panima Publishing Corporation, 4th Ed., 2003.
2. Glick, B. R and Pasternak, J. J. “Molecular Biotechnology-Principles and applications of Recombinant DNA”. ASM Press, Washington DC, 1994.
3. Nicholl, D. S. T. “An introduction to Genetic Engineering”. Cambridge University Press, 3rd Ed., 2008.
4. Nancy Craig et al. “Molecular Biology: Principles of Genome Function”, Oxford University Press, 1st Ed., 2010.
BIOPROCESS ENGINEERING
Subject Code : 14IBT14
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn the fundamental concepts of bioprocess engineering; To learn and understand fluid flow process, mixing process and mass transport; To apply the concepts of fluid flow, mixing and filtration to industrial operations; To understand and design measurement & control strategies for these operations.
Course Outcomes: At the end of this course, student will be able to:
· Demonstrate the concepts of fluid flow, mass transfer, mixing and filtration for industrial application.
· Identify rheological behavior and diffusion phenomena of fermentation broth.
· Apply mass transfer concepts to design aeration and agitation of fermentation process.
· Demonstrate knowledge of filtration and mixing process in industrial operation.
· Develop control strategies for bioprocess operations.
MODULE 1 INTRODUCTION 10 Hours
Introduction to bioprocess engineering, Balances: elemental balances, material balance (steady and unsteady) and heat balance, Energy balancing for bioreactors. Yield: The yield values for anaerobic & aerobic systems. Mass and energy yield coefficients, overall yield for microorganisms.
MODULE 2 FLUID FLOW AND MIXING 10 Hours
Fluid statics: Pressure at a point and measurement, osmotic pressure. Viscosity and its measurements: Newton’s laws of viscosity, Newtonian and non Newtonian fluids (NF &
NNF), Rheology of fermentation broth.
Fluid Flow: types of fluid flow, laminar, turbulent flow. Bernoulli equation. Flow measuring devices: Variable head & area meters, wheel flow meter. Hydrodynamic boundary layer, boundary layer shear force.
MODULE 3 MASS TRANSFER 10 Hours
Diffusion: Types of diffusion, Fick’s law, Role of diffusion in bioprocessing, L-L, L-S and G-L mass transfer.
Aeration: Oxygen uptake in cell culture, Gassed fluid, KLa and its measurement, oxygen supply and demand, sparger, aeration number, power requirement, bubble shear.
MODULE 4 UNIT OPERATIONS 10 Hours
Filtration: Filter aids, filtration theory. Centrifugation: centrifugation theory. Mixing: Mechanisms of mixing. Flow pattern: radial and axial flow impeller, mixing theory, mixing
time, effectiveness of mixing and power requirement.
MODULE 5 BIOPROCESS CONTROL 10 Hours
Concept of bioprocess control, Elements of feedback controller, types of controller action, advanced control strategies, controller tuning, online and offline measurements (P,T, pH, agitator speed, off gas analysis).
TEXT BOOKS:
1. Pauline M. Doran, “Bioprocess Engineering Principles”, Academic Press, 2nd Ed., 2012.
2. Stanbury, Vitaker and Hall, “Principles of Fermentation Technology”, Butterworth Heinemann, 2nd Ed., 1999.
REFERENCE BOOKS:
1. Shuler and Kargi, “Bioprocess Engineering”, PHI, 2nd Ed., 2001.
2. Brian McNeil and Linda Harvey (Ed), “Practical Fermentation Technology”, Wiley, 2008.
3. David Himmelblau, “Basic principles and Calculations in Chemical Engineering”, Prentice Hall. 6th Ed, 1996.
4. Donald R. Coughanowr, Lowell B. Koppel, “Process systems analysis and control”, MGH, 2nd Ed., 1991.
5. Richardson and Coulson, “Chemical Engineering”, Volume 1, Butterworth Heinemann, 6th Ed., 1999.
BIOPROCESS MODELING AND AUTOMATION
Subject Code : 14IBT151
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn the concepts and need for process modeling and simulation. To apply the concepts of modeling to linear and nonlinear bioprocesses. To apply the modeling principles to systems generating ordinary and partial differential model equations. To understand principle of stochastic modeling. To use and apply software tools for simulation of model equations.
Course Outcomes: At the end of this course, student will be able to:
· Understand the concepts and need for process modeling and simulation.
· Apply the concepts of modeling to linear and nonlinear bioprocesses.
· Apply the modeling principles to systems generating ordinary and partial differential model equations.
· Describe principle of stochastic modeling.
· Use and Apply software tools for simulation of model equations.
MODULE 1 PRINCIPLES OF MODELING 10 Hours
Concept of modeling and simulation, general aspects of modeling, dependent and independent variables, classification of models. Material and energy balance equations,
constitutive equations, general strategy of modeling, Solution strategies and simulation. Measurements, errors and accuracy. Modeling of simple systems.
MODULE 2 LINEAR AND NON LINEAR EQUATIONS 10 Hours
Elemental balances and degrees of reduction, extractor, absorber. Models of enzyme kinetics (Michaelis-Menten), growth kinetics (Monod) and product formation kinetics. Receptorligand dynamics, RT-PCR modeling. Numerical solutions to linear and nonlinear algebraic equations.
MODULE 3 ORDINARY DIFFERENTIAL EQUATIONS 10 Hours
Models of predator-prey, commensalism and mutualism, Structured kinetic models, pharmacokinetic models. Bioreactors modeling (MFR and PFR with linear and nonlinear
kinetics), models of heat transfer and mass transfer in bioreactor. Numerical solutions to ODEs.
MODULE 4 PARTIAL DIFFERENTIAL EQUATIONS & STOCHASTIC MODELING 10 Hours
Kinetics of immobilized system with internal mass transfer, diffusion across biological membranes, fluid flow in physiological vessel (blood flow), numerical solutions to PDEs. Principles of stochastic modeling, age distribution of microbial cells, budding of yeast cells.
MODULE 5 MODEL SIMULATION 10 Hours
MATLAB: Basic commands, plotting tools, matrices and operation, flow control, solving linear, nonlinear equations, ODEs, PDE toolbox, SIMULINK. Use of MATLAB to solve
problems formulated in Unit 1 to Unit 4.
TEXT / REFERENCE BOOKS
1. I.J. Dunn, E. Heinzle, J. Ingham and J.E. Prenosil, “Biological Reaction Engineering”, Wiley-VCH, 2nd Ed., 2003.
2. Stanley M. Dunn, Alkis Constantinides and Prabhas V. Moghe, “Numerical Methods in Biomedical Engineering”, Academic Press, 2006.
3. W. Fred Ramirez, “Computational Methods in Process Simulation”, Elsevier, 2nd Ed, 1998.
4. Ashim K. Datta, “Biological and Bioenvironmental Heat and Mass Transfer”, Marcel Deccer Inc., 2002.
5. Jens Nielsen, John Villadsen and Gunnar Liden, “Bioreaction Engineering Principles”, Plenum Publishers, 2nd Ed., 1994.
6. Pauline M. Doran, “Bioprocess Engineering Principles”, Academic Press, 2nd Ed., 2012.
INSTRUMENTAL METHODS OF ANALYSIS
Subject Code : 14IBT152
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn fundamentals of analytical methods; To understand various components of instrumentation system used in analysis; To learn the concepts and
applications of spectroscopic, chromatographic and electrophoretic techniques used for analysis of biomolecules; To understand working principle and instrumentation system of spectroscopic, chromatographic and electrophoretic techniques.
Course Outcomes: At the end of this course, student will be able to:
· Explain application of electromagnetic radiation in biomolecule analysis.
· Demonstrate fundamental concepts of analytical procedures like sampling, sample preparation, use of calibration of analytical methods, and Identify suitable technique.
· Explain the fundamental concepts and applications of spectroscopic, chromatographic and electrophoretic techniques.
· Understand working principle of instrumentation system of spectroscopy, chromatography and electrophoresis.
· Apply concepts of spectroscopic, chromatographic and electrophoretic techniques to analyse biomolecules qualitatively and quantitatively.
MODULE 1 INTRODUCTION 10 Hours
Introduction to analytical methods, types of analytical methods, selection of analytical method (accuracy, precision, sensitivity, selectivity, scale, time and cost).
Measurement and error: Types of error, measurement of error and accuracy.
Electromagnetic radiation: Properties of electromagnetic radiation, interaction of radiation with matter, Born – Oppenheimer approximation.
Sources of radiation: Continuous sources of UV, visible and IR radiation (D2, Tungsten filament, Xenon arc lamps, Nernst glower, Globar sources).
Components of an analytical instrument, signal amplifiers (Transistors, Operational Amplifiers), noise, signal to noise ratio, sources of noise, signal to noise improvement.
Sampling: types of samples, sample preparation, sample size, sampling error, stock solutions, sample dilution.
Calibration methods: reagent blank, one point calibration, linear calibration, standard addition method, internal and external standard.
MODULE 2 ABSORPTION & EMISSION SPECTROSCOPY 10 Hours
Optical spectroscopy: Source, optical components, wavelength selector, sample holders, detectors.
UV-Visible spectroscopy: Theory (Beer – Lambert’s law), chromophores and their characteristic absorption, theory of UV absorption (electronic transition – n to pi*, pi to pi*,
sigma to sigma*; Solvatochromism, Conjugated dienes – Woodward Fieser rules), instrumentation (single and double beam), qualitative and quantitative analysis, single and
multiple component analysis, numericals.
Infrared spectroscopy: Theory, instrumentation, qualitative analysis, FT-IR.
Atomic absorption spectroscopy: Theory, instrumentation and applications.
Fluorescence and Phosphorescence spectroscopy: Theory, instrumentation and applications.
MODULE 3 RESONANCE & SCATTERING SPECTROSCOPY 10 Hours
Nuclear magnetic resonance spectrometry: Theory (Larmor Equation), environmental effects on pNMR, chemical shift, spin-spin splitting, applications of pNMR, data interpretation.
Molecular mass spectrometry: Theory, methods of ionization (EI, ESI, Ion Spray, MALDI), mass analyzers (Magnetic sector, Quadrupole, TOF), MALDI-TOF in protein analysis and applications.
Turbidimetry: Theory, instrumentation and applications. Introduction to ICP-MS, ICP-OES.
MODULE 4 CHROMATOGRAPHIC TECHNIQUES 10 Hour
Introduction to chromatographic separations, classification. Basic principles and theory of chromatography (Plate theory, Rate theory – van Deemter equation), numericals. Gas chromatography and HPLC: principle, instrumentation, column, detector, mobile phase, sample preparation. Application of chromatographic techniques.
MODULE 5 ELECTROPHORETIC TECHINQUES 10 Hours
General principle, support media- Agarose gel, starch gel, agarose starch gel, polyacrylamide gel. Electrophoresis of protein: SDS-PAGE, native gels, gradient gels, isoelectric focusing gels, 2D polyacrylamide gel electrophoresis, cellulose acetate electrophoresis. Detection, estimation and recovery of proteins in gels. Electrophoresis of nucleic acids. Capillary electrophoresis: Zeta potential, Electro-endo-osmotic flow, Instrumentation, detectors, Applications.
TEXT / REFERENCE BOOKS:
1. Willard and Merit, “Instrumental Methods of Analysis”, CSS Publishers, 1986.
2. Douglas A. Skoog, F. James Holler and Timothy A. Nieman, “Principles of Instrumental Analysis”, Harcourt Brace College Publishers, 5th Ed., 1998.
3. R.M. Silverstein and W.P. Webster, “Spectrometric Identification of Organic Compounds”, Wiley & Sons, 7th Ed., 2005.
4. Chatwal & Anand, “Instrumental Methods of Chemical Analysis”, Himalaya Publishing House, 5th Ed., 2013.
5. K. Wilson and J. Walker, “Principles and Techniques of Practical Biochemistry”, Cambridge University Press, 1994.
6. S. Ahuja & N. Jespersen, “Modern Instrumental Analysis”, Elsevier, 2006
7. David Harvey, “Modern Analytical Chemistry”, MGH, 1st Ed., 2000.
8. B. Sivasankar, “Instrumental Methods of Analysis”, Oxford University Press, 2012.
NUMERICAL METHODS & BIOSTATISTICS
Subject Code : 14IBT11
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn numerical methods and statistical techniques to evaluate biological and bioprocess data. To understand the needs of statistical analysis in experimental design and analysis. To apply statistical methods for design of experiments and interpret the results. To apply statistical techniques to microarray analysis and genome mapping.
Course Outcomes: At the end of this course, student will be able to:
· Demonstrate methods for data representation using statistical tools.
· Apply statistical tools to biological problems and experimental designs.
· Understand the importance of statistics in solving biological problems.
· Perform ANOVA for biological data.
· Design and formulate statistical designs for experimentation.
· Evaluate and interpret the statistical analysis of biological data.
· Apply statistical techniques to genome mapping and interpret the outcomes.
MODULE 1: 10 Hours
Introduction to statistics and study design: Introduction to statistics, data, variables, types of data, tabular, graphical and pictorial representation of data. Significance of statistics to biological problems, experimental studies; randomized controlled studies, historically controlled studies, cross over, factorial design, cluster design, randomized; complete, block, stratified design, biases, analysis and interpretation.
MODULE 2: 10 Hours
Descriptive statistics and Observational study design: Types of variables, measure of spread, logarithmic transformations, multivariate data. Basics of study design, cohort studies, case-control studies, outcomes, odd ratio and relative risks.
Principles of statistical inference: Parameter estimation, hypothesis testing. Statistical inference on categorical variables; categorical data, binomial distribution, normal distribution, sample size estimation.
MODULE 3: 10 Hours
Comparison of means: Test statistics; t-test, F distribution, independent and dependent sample comparison, Wilcoxon Signed Rank Test, Wilcoxon-Mann-Whitney Test, ANOVA.
Correlation and simple linear regression: Introduction, Karl Pearson correlation coefficient, Spearman Rank correlation coefficient, simple linear regression, regression model fit, inferences from the regression model, ANOVA tables for regression.
Multiple linear regression and linear models: Introduction, Multiple linear regression model, ANOVA table for multiple linear regression model, assessing model fit, polynomials and interactions. One-way and Two-way ANOVA tables, F-tests. Algorithm and implementation using numerical methods with case studies.
MODULE 4: 10 Hours
Design and analysis of experiments: Random block design, multiple sources of variation, correlated data and random effects regression, model fitting. Completely randomized design, stratified design. Biological study designs. Optimization strategies with case studies.
MODULE 5: 10 Hours
Statistics in microarray, genome mapping and bioinformatics: Types of microarray, objectives of the study, experimental designs for micro array studies, microarray analysis, interpretation, validation and microarray informatics. Genome mapping, discrete sequence matching, programs for mapping sequences with case studies.
TEXT / REFERENCE BOOKS:
1. Alvin E. Lewis, “Biostatistics”, McGraw-Hill Professional Publishing, 2013.
2. J.D. Lee and T.D. Lee. “Statistics and Numerical Methods in BASIC for Biologists”, Van Nostrand Reinhold Company, 1982.
3. Wolfgang Boehm and Hartmut Prautzsch. “Numerical Methods”, A K Peters/CRC Press, 1993.
4. T.P. Chapman & Hall (Ed.). “Statistical Analysis of Gene Expression Microarray Data”, CRC Press, 2003.
5. G. Gibson & S.V. Muse. “A Primer of Genome Science”, Sinauer Associates, 2001.
6. R. Gentleman (Ed.). “Bioinformatics and Computational Biology Solutions using R and Bioconductor”, Springer, 2005.
7. John F. Monahan. “Numerical Methods of Statistics” (Cambridge Series in Statistical and Probabilistic Mathematics), Cambridge University Press Edition, 2011.
8. Joe D. Hoffman. “Numerical Methods for Engineers and Scientists”, CRC Press, 2nd Edition, 2001.
9. Warren J. Ewens Gregory Grant. “Statistical Methods in Bioinformatics: An Introduction” (Statistics for Biology and Health), Springer, 2005.
10. T. Hastie, R. Tibshirani, J. H. Friedman. “The Elements of Statistical Learning” Springer, 2001.
11. Warren John Ewens, Gregory R. Grant, Gregory Grant, R. “Statistical Methods in Bioinformatics”, Springer, 2005.
12. W.H. Press et al. “Numerical recipes- The Art of Scientific Computing”, Cambridge University, 3rd Ed., 2007.
BIOREACTION ENGINEERING
Subject Code : 14IBT154
IA Marks : 50
No. of Lecture Hrs./ Week : 04 Exam Hrs : 03
Total No. of Lecture Hrs. : 50 Exam Marks : 100
Course Objectives: To learn kinetics of enzymatic reactions and to understand enzyme substrate models of enzyme reactions; To analyse the effects of parameters affecting enzyme kinetics and to identify and formulate methods to evaluate enzyme kinetics in homogeneous and heterogeneous systems; To analyse mass transfer effects on enzyme kinetics and to know the technologies of production of industrial enzymes; To learn and understand methods of protein purification for applications at higher concentrations.
Course Outcomes: At the end of this course, student will be able to:
· Explain enzyme substrate models and kinetics of enzyme reaction.
· Demonstrate effects of process parameters on enzyme reactions.
· Formulate evaluation methods for kinetic parameters for homogeneous and heterogeneous enzyme reactions.
· Analyse mass transfer effects involved in immobilized enzyme systems.
· Explain production of industrial enzymes.
· Describe protein enrichment or purification methods.
MOULE 1 BIOLOGICAL KINETICS 10 Hour
Enzyme nomenclature and enzyme classification, energy potentials of enzyme (Stern layers).
Types of reaction, elementary and non elementary reaction, molecularity and order of reaction, The enzyme-substrate complex and enzyme action, simple enzyme kinetics with one and two substrate. Derivation of Michaelis-Menten kinetics, Briggs-Haldane approach, Monod equation. Double Michaelis–Menten kinetics, allosteric kinetics, effect of temperature and pH on enzyme kinetics. Substrate and product inhibition of growth. Substrate uptake kinetics. Interacting microorganisms.
MOULE 2 ENZYME REACTION IN HOMOGENEOUS SYSTEMS 10 Hour
Basic reaction theory, reaction thermodynamics, Reaction rate & kinetics: first, second and zero order reaction. Estimation of reaction rate: integral & differential method. Enzyme kinetics: Michaelis-Menten Kinetics Enzyme immobilization kinetics, enzyme deactivation kinetics, Mass transfer limitations. Enzyme inhibition kinetics (substrate, product, inhibitor), Competitive, Noncompetitive and Mixed Inhibition kinetics.
MOULE 3 ENZYME REACTION IN HETEROGENEOUS SYSTEMS 10 Hour
Catalyst immobilization, substrate concentration profile in an immobilized biocatalyst particle. Steady state shell balance. Zero, first order and M-M kinetics. Concentration profile in other geometry. Dimensionless parameters from diffusion reaction model. Effect of internal and external mass transfer. Effect of Mass-Transfer Resistance.
MOULE 4 INDUSTRIAL ENZYMES & APPLICATIONS 10 Hour
Enzyme engineered for new reactions-novel catalyst for organic synthesis. Case studies: thermozymes cold adopted enzymes. Ribozymes, hybrid enzymes, diagnostic enzymes, therapeutic, inteins. enzymes of industrial importance (amylase, glucose isomerase, cellulose, lipase, protease, xylanase, invertase, peroxidases).
MOULE 5 ENZYME PURIFICATION 10 Hour
Separation of insolubles: filtration, centrifugation. Extraction and purification of solubles: Ultra filtration, high performance tangential flow filtration, Liquid liquid extraction (ATPS). Recovery and purification of intracellular products: cell disruption, chromatographic techniques. Analytical assays of purity level of enzymes.
TEXT BOOKS:
1. Pauline M. Doran, “Bioprocess Engineering Principles”, Academic Press, 2nd Ed., 2012.
2. El-Mansi (Ed.), “Fermentation Microbiology and Biotechnology”, CRC Press, 3rd Ed., 2011.
REFERENCE BOOKS:
1. Ashok Pandey et al., “Enzyme Technology”, Springer Publisher, 2006.
2. Nielsen et al., “Bioreaction Engineering Principles”, Plenum Publishers, 2nd Ed., 2002.
3. Mohammed A. Desai (Ed.), “Downstream Processing of Proteins: Methods and Protocols”, Humana Press, 2000.
4. Satinder Ahuja, “Handbook of Bioseparations”, Vol 2, Academic Press, 1st Ed., 2000.
5. Devasena, T. “Enzymology”, Oxford University Press, 2012.
6. Marangoni, “Enzyme kinetics: A modern approach”, Wiley India 2012.
FERMENTATION TECHNOLOGY & MOLECULAR BIOLOGY LAB
Subject Code : 14IBT16
IA Marks : 25
No. of Practical Hrs./ Week : 03 Exam Hrs : 03
Total No. of Practical Hrs. : 36 Exam Marks : 50
Course Objectives: To learn the methods involved in preparation of medium for microbial and plant cell culture. To understand methods of medium design and reduction of lag phase. To gain hands on experience in plant tissue culture and molecular biology techniques.
Course Outcomes: At the end of this course, student will be able to:
· Prepare and develop inoculum for industrial fermentation.
· Design medium for optimal fermentation and reduce lag period.
· Design and Perform experiments on plant tissue culture.
· Design and Perform molecular biology experiments.
EXPERIMENTS
1. Preparation of inoculums and aseptic inoculum transfer into media.
2. Preparation of medium for microbial culture, Media optimization using RSM.
3. Study of growth kinetics using different carbon sources.
4. Strategy to reduce lag phase.
Case Inoculum media Production media
a. Same composition Same composition
b. Different Composition Different composition
[Compare growth curve of case 1 & 2 using same microorganism]
5. Production of callus, preparation of media and suspension culture.
6. Secondary metabolite production using suspension culture.
7. Plasmid Isolation by Mini Prep Method.
8. Restriction Digestion and Restriction Mapping Technique.
9. PCR Technique and the Use of Gel-Doc System.
10. Salt Extraction and Estimation of High Quality Genomic DNA obtained from Plant Source.
11. Small-Scale Extraction and Estimation of RNA obtained from Plant Source.
12. Western blotting technique.