DSP Theory Course File
DSP Theory Course File
2
LECTURE PLAN
Unit I
Course Outcomes:
C402.1:
Students understand fundamentals of downstream processing for product recovery, unit
operations involved in bioseparation and understand the various methods of cell disruption.
Course Content:
UNIT I Downstream Processing
Introduction to downstream processing principles characteristics of biomolecules and
bioprocesses. Cell disruption for product release – mechanical, enzymatic and chemical methods.
Pretreatment and stabilisation of bioproducts.
Content delivery methods:
Black Board Power Point Presentation Student Assignment
Student Seminar Add-on Courses - Live Demonstration -
NPTEL Videos Any Other Methods
Assessment methods:
Assignments Tests / Model Exams Univ. Exams
Lab Practices - Seminar/Student Viva Mini / Major Projects -
Add-on Courses - Certifications - Others -
Content delivery methods:
Knowledge Delivery No. of hrs to
Sl. No. Course Content CO Statement
level method be handled
Chalk and
1. Introduction to downstream processing. C402.1 U Board
1
Role, importance and principles Chalk and
2. C402.1 U Board
2
downstream processing
Characteristics of microorganisms,
biomolecules, fermentation broth and Chalk and
3. C402.1 U/An Board 1
other factors affecting downstream
processing
Types of bioproducts,
Chalk and
4. Purification of intracellular products C402.1 U Board 2
Cell disruption methods - Classification
Mechanical and Physical cell disruption Chalk and
5. C402.1 U Board/PPTs 1
methods
Enzymatic and chemical cell disruption Chalk and
6. C402.1 U Board/PPTs 1
methods
Pretreatment and stabilisation of Chalk and
7. C402.1 U/An Board 1
bioproducts.
Chalk and
8. Purification of Recombinant Proteins C402.1 U/An Board/PPTs
1
R- Remembering; U- Understanding; Ap- Applying; An- Analysing; E- Evaluating; C- Creating
References:
1. Bioseparations by Belter – page nos.
2. Bioseparations by B. Sivasankar – page nos.
3. Class notes / Portal notes/NPTEL videos
3
LECTURE PLAN
Unit II
Course Outcomes:
C402.2:
Students gain the ability to design filtration and centrifugation operation for separation of
biomass.
Course Content:
UNIT II Physical Methods of Separation
Unit operations for solid-liquid separation - filtration and centrifugation.
Content delivery methods:
Black Board Power Point Presentation Student Assignment
Student Seminar Add-on Courses - Live Demonstration -
NPTEL Videos Any Other Methods
Assessment methods:
Assignments Tests / Model Exams Univ. Exams
Lab Practices - Seminar/Student Viva Mini / Major Projects -
Add-on Courses - Certifications - Others -
Content delivery methods:
Knowledg Delivery No. of hrs to
Sl. No. Course Content CO Statement
e level method be handled
Solid – liquid separation, Various
1. Chalk and
methods available and their role in C402.2 U/An Board
1
downstream processing
Principles of sedimentation and settling
2. Chalk and
in solid liquid separation and their C402.2 U Board
1
demerits
3. Filtration operation – Principles and Chalk and
C402.2 U/An Board 1
applications
4. Classification of filtration methods Chalk and
C402.2 R/U Board/PPTs 2
Filtration equipments (batch and continuous)
Derivation for constant rate and
5. Chalk and
constant pressure filtrations and C402.2 R/Ap Board
2
problem solving
6. Centrifugal separation – Principles and Chalk and
C402.2 R/U Board/PPTs
1
types
Chalk and
7. Various equipments for centrifugal
C402.2 U/An Board/ NPTEL 2
separation Videos
8. Scale-up of centrifugal separations & Chalk and
C402.2 U/An Board 2
Problem solving
R- Remembering; U- Understanding; Ap- Applying; An- Analysing; E- Evaluating; C- Creating
References:
1. Bioseparations by Belter – page nos.
2. Bioseparations by B. Sivasankar – page nos.
3. Class notes / Portal notes/NPTEL videos
4
LECTURE PLAN
Unit III
Course Outcomes:
C402.3:
Students learned to identify a suitable unit operation for isolation and concentration for the
given bioproduct.
Course Content:
UNIT III Isolation of Products
Adsorption, liquid-liquid extraction, aqueous two-phase extraction, membrane separation –
ultrafiltration and reverse osmosis, dialysis, precipitation of proteins by different methods.
Content delivery methods:
Black Board Power Point Presentation Student Assignment
Student Seminar Add-on Courses - Live Demonstration -
NPTEL Videos Any Other Methods
Assessment methods:
Assignments Tests / Model Exams Univ. Exams
Lab Practices - Seminar/Student Viva Mini / Major Projects -
Add-on Courses - Certifications - Others -
Content delivery methods:
Knowledg Delivery No. of hrs to
Sl. No. Course Content CO Statement
e level method be handled
Products isolation - Various methods
1. Chalk and
available and their importance in C402.3 U/An Board
1
downstream processing
Chalk and
2. Principles of adsorption operation (batch
C402.3 U Board/ NPTEL 3
and continuous) and Problem solving Videos
Chalk and
3. Principles of liquid-liquid extraction
C402.3 U/An Board/ NPTEL 3
(batch and continuous) and Problem solving Videos
4. Principles of aqueous two-phase Chalk and
C402.3 R/U Board/PPTs
1
extraction and applications
5. Membrane separation – Principles, Chalk and
C402.3 R/U Board 1
classification.
Operation and applications of
6. Chalk and
ultrafiltration and reverse osmosis, C402.3 R/An/Ap Board/PPTs 1
dialysis
7. Proteins precipitation – by solvent Chalk and
C402.3 U/An Ap Board 1
modification methods
8. Proteins precipitation – by solute Chalk and
C402.3 U/An/Ap Board 1
modification methods
R- Remembering; U- Understanding; Ap- Applying; An- Analysing; E- Evaluating; C- Creating
References:
1. Bioseparations by Belter – page nos.
2. Bioseparations by B. Sivasankar – page nos.
3. Class notes / Portal notes/NPTEL videos
5
LECTURE PLAN
Unit IV
Course Outcomes:
C402.4:
Students should be able to select a suitable chromatographic operation for purification of given
bioproducts.
Course Content:
UNIT IV PRODUCT PURIFICATION
Chromatography – principles, instruments and practice, adsorption, reverse phase, ion-exchange,
size exclusion, hydrophobic interaction, bioaffinity and pseudo affinity chromatographic
techniques.
Content delivery methods:
Black Board Power Point Presentation Student Assignment
Student Seminar Add-on Courses - Live Demonstration -
NPTEL Videos Any Other Methods
Assessment methods:
Assignments Tests / Model Exams Univ. Exams
Lab Practices - Seminar/Student Viva Mini / Major Projects -
Add-on Courses - Certifications - Others -
Content delivery methods:
Knowledge Delivery No. of hrs to
Sl. No. Course Content CO Statement level method be handled
Principles, mechanism and
1. Chalk and
classification of chromatography. Role C402.4 U/An Board 2
of chromatography in bioseparation
2. Various steps involved in Chalk and
C402.4 U Board/PPTs
1
chromatography and Instrumentation
3. Working principle and applications of Chalk and
C402.4 U/An Board/PPTs 1
adsorption chromatography
Working principle and applications of
4. Chalk and
size exclusion and reverse phase C402.4 R/U Board/PPTs 2
chromatography
5. Working principle and applications of Chalk and
C402.4 R/Ap Board/PPTs 1
ion-exchange chromatography
Working principle and applications of
6. Chalk and
hydrophobic interaction C402.4 R/U Board/PPTs 1
chromatography
7. Working principle and applications of Chalk and
C402.4 U/An Board/PPTs 2
bio and pseudo affinity chromatography
8. Design and scale-up of Chalk and
C402.4 U/An Board 2
chromatography & Problem solving
R- Remembering; U- Understanding; Ap- Applying; An- Analysing; E- Evaluating; C- Creating
References:
1. Bioseparations by Belter – page nos.
2. Bioseparations by B. Sivasankar – page nos.
3. Class notes / Portal notes/NPTEL videos
6
LECTURE PLAN
Unit V
Course Outcomes:
C402.5:
Students understand the importance of final polishing of bioproducts and applications of various
methods.
Course Content:
UNIT V FINAL PRODUCT FORMULATION AND FINISHING OPERATIONS 8
Crystallization, drying and lyophilization in final product formulation.
Content delivery methods:
Black Board Power Point Presentation Student Assignment
Student Seminar Add-on Courses - Live Demonstration -
NPTEL Videos Any Other Methods
Assessment methods:
Assignments Tests / Model Exams Univ. Exams
Lab Practices - Seminar/Student Viva Mini / Major Projects -
Add-on Courses - Certifications - Others -
Content delivery methods:
Knowledge Delivery No. of hrs to
Sl. No. Course Content CO Statement level method be handled
1. Significance of product formulation Chalk and
C402.5 U/An Board 1
and finishing operations for products
2. Various methods available for product Chalk and
C402.5 R/U Board
1
formulation and finishing operations
3. Crystallization – Principles and types. Chalk and
C402.5 R/U Board 1
Solid – liquid equilibrium.
4. Supersaturation methods Chalk and
C402.5 R/U Board/PPTs 1
Nucleation - types Nucleation
5. Crystallization equipments (batch and Chalk and
C402.5 U/Ap Board
1
continuous)
6. Principle, operation and applications of Chalk and
C402.5 R/U/Ap Board/PPTs
1
freeze drying
7. Principle of drying, types of drying and Chalk and
C402.5 U/An Board
2
drying equipments
8. Chalk and
Rate of drying & Problem solving C402.5 U/Ap/An Board
2
R- Remembering; U- Understanding; Ap- Applying; An- Analysing; E- Evaluating; C- Creating
References:
1. Bioseparations by Belter – page nos.
2. Bioseparations by B. Sivasankar – page nos.
3. Class notes / Portal notes/NPTEL videos
7
St. Joseph’s College of Engineering,
Department Of Biotechnology
Model I
Branch : Biotechnology Subject : Downstream Processing
Semester : VII Subject Code : BT6702
Part – A (10 x 2 = 20 marks)
Sl. No. Question Knowledge COs
level
1. Define thermolysis. R C402.1
2. Name some physical methods of cell disruption. R/U C402.1
3. What are the factors that impact difficulty and cost of recovery C402.1
An/E
of bioproducts?
4. What are the parameters influence the rate and degree of cell C402.1
An
disruption in high pressure homogenizer?
5. What is the mechanism of cell disruption by alkali treatment? R/U C402.1
6. Define rate of filtration. R/U C402.2
7. Name the unit operations used for the separation of suspended C402.2
R
solids.
8. What are the pretreatment methods used before filtration? R/U C402.2
9. What is centrifugal filtration? R/U C402.2
10. When centrifugal separators preferred than filtration? An/Ap C402.2
Part – B (5 x 16 = 80 marks)
11. (a) Explain the various chemical methods of cell disruption. (OR) R/U C402.1
(b) Discuss in detail on various characteristics of fermentation broth C402.1
U/An
that influence the separation of biomolecules.
13. (a) Write a detailed account on choice of cell disruption methods. (OR) U/An C402.1
(b) Explain the basic steps and unit operations involved in downstream C402.1
R/U
processing.
15. (a) Explain the various cross cutting strategies adopted by C402.1
U/Ap/An
bioseparation. (OR)
(b) What is constant pressure filtration? Give examples. Derive the C402.2
R/U/Ap
expression for rate of filtration in constant pressure filtration.
17. (a) Explain the working of continuous filtration equipment with neat C402.2
R/U
sketch. (OR)
(b) Explain the principle and operation of disc stack centrifuge and C402.2
R/U/Ap
derive the equation for volumetric capacity.
19. (a) Explain the principle and operation of tubular bowl centrifuge and R/U/Ap C402.2
derive the equation for volumetric capacity. (OR)
(b) A tubular bowl centrifuge is used to recover yeast cells from fermentation U/Ap/An C402.2
broth. At the flow rate of 10 lit/min and 5000 rpm 50% of the cells can be
recovered. You are now asked by your manager to imcrease the recovery to
90% using the same equipment.
i) What flow rate should you use?
ii) If you double the rotation speed how much does the
centrifugation velocity change.
8
St. Joseph’s College of Engineering,
Department Of Biotechnology
Model II
Part – B (5 x 16 = 80 marks)
11. (a) Discuss the principle and practice of various batch adsorption C402.3
in bioseperation. (OR) R/U
(b) The partition coefficient value of an organic acid in solvent C402.3
system is 2.7
i. Calculate the volume of organic solvent required to extract
U/AP/An
99% of the acid from 50 ml of aqueous solution.
ii. How many extractions with 50ml of the organic solvent
would be required to extract 99% of the acid.
13. (a) Discuss the solvent precipitation of bioproducts with their C402.3
R/U/An
merits and demerits. (OR)
(b) Explain the method of aqueous two phase extraction in detail R/U C402.3
15. (a) Explain the process of protein precipitation using ammonium C402.3
U/Ap
sulphate. (OR)
(b) Give a detailed account on gel permeation chromatography. U/Ap C402.4
9
17. (a) Explain the factors affecting chromatography column C402.4
efficiency
U/An
Discuss the important parameters to be considered in selection of
chromatography separation process. (OR)
(b) Describe in detail pseudo affinity chromatography separation C402.4
U/Ap/An
process with application in bioseparation.
19. (a) Describe in detail about bioaffinity chromatography separation U/Ap/An C402.4
process with application in bioseparation. (OR)
(b) Derive and discuss the equation for determination of column U/An C402.4
height in chromatography.
Assignment I
Part-A
Sl. No. Question Knowledge COs
level
1. What are the characteristics of fermentation broths? R/U C402.1
2. What are the different stages in bioseparation? R/U C402.1
3. Name some the physical methods of cell disruption? R C402.1
4. How will you prevent the thermal denaturation of C402.1
An/Ap
products during ultrasoniction?
5. Write short notes on alkali treatment? R/U C402.1
6. What is the principle of detergent solubilization? R/U C402.1
7. Define cell wall permeabilization. R/U C402.1
8. What are the different methods of Mechanical disruption C402.1
R/U
of cells?
Part – B
9. Explain the various chemical methods of cell disruption. R/U C402.1
10. Explain the working principle of High pressure C402.1
U/An
homogenizer. Write their merits and demerits.
11. Describe the role and importance of downstream C402.1
U/An
processing in bioprocess.
12. Write a detailed essay on characteristics of fermentation C402.1
U/An
broth that influence the separation of biomolecules
10
St. Joseph’s College of Engineering,
Department Of Biotechnology
Assignment II
Part-A
Part – B
9. Discuss the various methods of scale up of centrifuges. R/U C402.2
10. Explain with neat sketch the working principle of C402.2
R/U/
continuous filtration equipment
11. You plan to concentrate an E.coli cell suspension by C402.2
centrifugation prior to cell disruption. The suspension has
the following physical properties. Cell dia. 1 μm, density
difference 50 kg/m3, viscosity 2*10-3 kg/m sec, the
centrifuge length is 13.5 inch and the centrifuge diameter
is 5 inch.
(i) If the maximum allowable cake thickness
in the bowl is 1.5 inch, predict the
throughput (Volumetric capacity) for the
proposed operation. Assume the centrifuge U/An/Ap
will be operated at 16,000 rpm and that it
will deliver 90% of the theoretical capacity.
(ii) After cell disruption the diameter of the
cell debris has been reduced to an average of
0.5 μm and the viscosity of the suspension
has been increased to 8*10-3 kg/m sec.
Estimate the new capacity of the centrifuge
for this operation. Assume the centrifuge
operates at the same speed and efficiency.
12. Explain the principle and operation of Disc stack C402.2
centrifuge and derive the equation for volumetric U/An
capacity.
11
St. Joseph’s College of Engineering,
Department Of Biotechnology
Assignment III
Part-A
Part – B
9. Write a short essay on (i) Salting-out (ii) Microfiltration C402.3
R/U/
10. Explain the method of aqueous two phase extraction in C402.3
R/U/
detail.
11. We have measured the adsorption of the antibiotic C402.3
(gentamicin) from a liquid at pH 9.5 onto a non-ionic
carbon. We find the following results.
12
organic solvent required to extract 99% of the acid from
50 ml of aqueous solution. ii) How many extractions with
50 ml of the organic solvent would be required to extract
99% of the acid?
13
St. Joseph’s College of Engineering,
Department Of Biotechnology
Assignment IV
Part-A
Part – B
9. Discuss the principle of separation of charged species by C402.4
ion exchange chromatography in detail with application U/An
in bioseparation.
10. Describe in detail about bioaffinity chromatographic C402.4
U/An
separation process with application in bioseparation.
11. Derive and discuss the equation for determination of C402.4
U/An
column height in chromatography.
12. Explain the factors affecting chromatography column C402.4
U/An
efficiency.
14
St. Joseph’s College of Engineering,
Department Of Biotechnology
Assignment V
Part-A
Part – B
15
Attainment of COs – Based on Average Attainment
16