Recombinant DNA > Sing 2 > Flashcards

Sing 2 Flashcards

(53 cards)

Study These Flashcards
1
Q

What are the 2 types of single use bioreactors?

A
  1. stirrer type
  2. rocking motion type
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What parts does a typical bioreactor consist of?

A
  1. Agitator
  2. Baffle
  3. Sparger
  4. Jacket
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

What is the use of an Agitator in a bioreactor?

A

mixing of the contents of the reactor

–> keeps cells in perfect homogenous condition for better transport of nutrients & oxygen to desired product

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What is the use of a Baffle in a bioreactor?

A

break the vortex formation in the vessel, consumes additional power

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What is the use of a Sparger in a bioreactor?

A

In aerobic cultivation process
–> supply adequate oxygen to the growing cells

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What is the use of a Jacket in a bioreactor?

A

provides circular area for circulation of diff temp of water
–> maintain temp of bioreactor

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What are the basic functions of a bioreactor?

A
  1. provide controlled environment for optimum product yields
  2. permit aseptic fermentation & meet requirements of containment regulations eg prevention of escape of viable cells into environment
  3. provide adequate mixing and aeration for optimum growth
  4. minimum power consumption
  5. easy & dependable temp control
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What are the various types of fermenters for biotech?

A
  • Bacteria vs Mammalian or insect cells
  • Aerobic
  • Anaerobic
  • Stainless steel
  • Disposable bioreactor
  • Stirrer
  • Rocking motion
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What are the 2 types of cell cultures?

A

suspension and adhesive cell culture

adhesive cell culture require cleavage of anchorage proteins to release cells

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What prokaryote organism is preferred for the production of rProteins?

A

E. coli

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What are the advantages of using E coli for production of recombinant proteins?

A
  • high growth rate
  • simple media
  • ease of genetic manipulation
  • high yield
  • low cost
  • well-characerized genetics
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

What are the disadvantages of using E coli for production of recombinant proteins?

A
  • inclusion bodies
  • lack of post-translational modifications
  • endotoxin
  • poor secretion
  • antibiotic resistance gene
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Optimization of growth of microorganism in a particular media is desirable due to economical and availability of particular growth constituent.

True or False

A

True

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

What is the most common mode of bacterial cell division for E coli?

A

Binary division

single bacteria cell grows transversely with synthesis of chromosomal DNA.

tansverse septum appears in middle of cell body and divides cel into 2

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Name 2 methods for measuring bacterial growth in liquid media and in solid support media

A
  1. plate count method
  2. turbidimetric method
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Explain the principle of the plate count method.

A

bacterial culture suspension introduced onto solid support media –> grow colonies –> count no. of colonies with colony counter with lamp at bottom, grid and magnifying glass

No. of bacteria per ml = no of colonies counted on plate X dilution of sample

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

For measuring bacterial growth using plate count method, plate with colony
count of _________ can be used to determine the number of bacteria present in
original stock.

A

30-300

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

Explain the principle of the turbidimetric method

A

bacteria cell suspension placed in test cuvette w/ corresponding media in ref cuvette –> optical density/absorbance at 600nm measurd

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

What principle is turbidimetric method for cell counting based on?

A

light scattering principles of particulate matter

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

What is the disadvantage of turbidimetric method to measure cell number in suspension?

A

This method can not distinguish between live or dead bacteria as both form contribute to the turbidity.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

What are the 4 phases of growth of bacteria?

A
  1. lag phase
  2. exponential/log phase
  3. stationary phase
  4. death phase
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

What happens in the lag phase?

Study These Flashcards
A
  • bacteria gets adjusted to the new media and grow in size instead of dividing into daughter cells
  • synthesize crucial enzymes for optimal grwoth
  • cell is metabolically active
  • synthesize protoplasm
  • at end of phase, bacterai divides and enter next stage
23
Q

What happens in the log phase?

Study These Flashcards
A

-active division
-cell population is more or less
homogenous in terms of chemical composition, physiology & metabolic activity
-plot of number of cell (in log scale) against time gives straight line
- growth of bacterial cell
population increasing at constant rate & continues until substrate concentration is limiting

24
Q

What happens at the stationary phase?

Study These Flashcards
A

substrate limiting –> formation of inhibitory product such as organic acid –> log phase of growth decline –> no of cells constant as no. of division = no. of death

25
How does the cell population remain constant at stationary phase even though substrate is limiting and growth declines?
death of old cell provides enough nutrient for remaining cells to grow and multiply to maintain the constant number
26
What happens at the death phase?
substrate from dying cells not sufficient & waste produced --> death rate of bacteria higher than growth rate --> bacteria decline sharply
27
What are the factors that affect bacterial growth?
1. Nutrient availability (C, N, salts, ions, trace elements) 2. Temperature 3. mositure 4. pH 5. oxygen level 6. presence of inhibitors or antimicrobial agents 7. anti-foam
28
What factors influence bacterial death?
- heat - pH extreme - radiation - chemical agents
29
What are some mechanisms of bacterial cell death?
DNA damage, protein denaturation, and membrane disruption
30
What are the 3 culture feeding strategies?
1. batch 2. fed-batch 3. continuos
31
What is the batch strategy?
no extra feeding is used from beginning to end of the process
32
What is the fed-batch feeding strategy?
feeding w/ substrate & supplements can extend duration of culture for higher cell densities or switch metabolism to produce e.g. a recombinant protein
33
What is continuous culture strategy?
fresh medium is continuously added, while culture liquid containing left over nutrients, metabolic end products and microorganisms is continuously removed at the same rate to keep the culture volume constant
34
What are the 2 types of continuous culture systems?
1. Turbidostat --> growth rate controlled by maintaining constant cell density (turbidity) 2. Chemostat --> growth rate of microorganisms is controlled by limiting the supply of one or more essential nutrients (e.g., C, N, P)
35
What are the parameters to be optimized in fermentation?
* Medium * Temperature * Oxygen * pH * Pressure * Nine mineral salts * (Codon optimization) * Different strains
36
What are the nine mineral salts that need to be optimized in fermentation?
ammonium, potassium and sodium cations; and carbonate, chloride, nitrate and sulphate anions.
37
What are the different types of medium?
1. defined medium 2. complex medium 3. serum and animal componenet-free medium
38
What is a defined medium?
specific chemicals at known concentrations
39
What is a complex medium?
mixtures of yeast extract or enzymatic digests of protein (e.g. Casein hydrolysate); the exact amount and kinds of nutrients present in the medium are not known
40
What are the problems associated with serum and animal component-free medium?
batch to batch variation, virus contamination, regulatory advantages
41
What is the composition of medium?
- water - carbon source - nitrogen source - trace elements - growth factors - nine mineral salts
42
How should the feeding strategy be designed?
so that the growth rate is maintained to limit the production of toxic formate, acetate and other metabolic compounds, enhancing bacterial growth.
43
What is the DO-stat method?
ses dissolved oxygen (DO) levels in a bioreactor to regulate the feeding of nutrients during microbial or cell culture growth. --> often used in fed-batch
44
What is Plackett-Burman Design (PBD)?
Experimental design used for screening experiments, where the goal is to identify the most significant factors (variables) from a larger set of factors that influence a process or response.
45
What is Response Surface Methodology (RSM)?
collection of mathematical and statistical techniques used to model and optimize processes where a response (output) is influenced by multiple variables (inputs). --> find optimal conditions
46
Name some experimental designs/methodologies used to optimize fermentation condions
- single factor analysis - Plackett-Burman Design (PBD) - Response Surface Methodology (RSM) - box-blenken design
47
What is single-factor analysis?
experimental approach where the effect of one independent variable (factor) on a response (dependent variable) is studied, while keeping all other factors constant. This method is commonly used in the early stages of experimentation to identify how a single variable influences the output.
48
What are the major steps invovled in a new drug development process?
1. target discovery 2. drug candidate discovery 3. preclinical testing 4. investigational new drug (IND) application 5. clinical trials 6. new drug application (NDA) 7. regulatory review 8. Approval 9. post-marketing (Phase 4) activities
49
What do preclinical testing assess?
drug absorption, distribution, metabolism, and excretion (ADME)
50
What shall IND application include?
drug's composition, preclinical data, proposed clinical trial design, and manufacturing details
51
What shall NDA application include?
clinical trial data, including efficacy, safety, and manufacturing information
52
What characteristics should a drug candidate be evaluated on?
efficacy, selectivity, and safety
53
Describe the process of drug candidate discovery
Use various mehods, including high-throughput screening and computer-aided drug design, to identify potential drug candidates that can interact with the target
Recombinant DNA
flashcards
Decks in class (9)
# Cards
Brainscape helps you reach your goals faster, through stronger study habits.
© 2026 Bold Learning Solutions. Terms and Conditions