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PHAR 124 > Potential Questions For Final > Flashcards

Potential Questions For Final Flashcards

1
Q

What are the methods of transformation?

A

Heat shock
CaCl2 transformation
Lipofectin and similar molecules
Electroporation
Microinjection

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2
Q

To make the recombinant plasmid permeable to DNA molecules which of the chemicals is added?

A

CaCl2

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3
Q

Generally a plasmid vector contains how many elements?

A

3

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4
Q

Which of the following enzymes is required for end to end joining of DNA?

A

DNA ligase

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5
Q

Which of the following is responsible for making a DNA copy from RNA?

A

Reverse transcriptase

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6
Q

Can two DNAs cut with different restriction enzymes join together to form a recombinant plasmid?

A

Yes, provided the two enzymes have the same reaction site

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7
Q

What are the differences between small molecule drugs and biological drugs?

A

Biologicals:
Bigger, higher molecular weight
Complex
Many options for modification
Produced in living cell culture
Can’t be characterized completely
Unstable
Immunogenic

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8
Q

What was the first biologic product?

A

Insulin

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9
Q

When do you extract primary metabolites and secondary metabolites?

A

Primary = log phase
Secondary = late log phase and early stationary phase

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10
Q

What are secondary metabolites?

A

Antibiotics and phenolics

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11
Q

At what level does rifamycin B work at?

A

Transcription level

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12
Q

What is hayflick’s phenomenon?

A

The number of passages decreases when cells are harvested from older individuals

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13
Q

What do most cells require as a supplement to promote cellular multiplication?

A

5-10% serum

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14
Q

Why choose CHO cells?

A

Good safety profile
Produce proteins with complex bio active PTMs that are similar to those produced in humans
Can grow in suspension culture
Can grow in serum-free chemically defined media
Allow gene amplification
Stronger expression units
Highly tolerant to pH, exogenous level, pressure, or temp

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15
Q

What is atryn?

A

Anti blood clotting protein antithrombin made using goats
Approved in 2009 by FDA

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16
Q

What was the first animal used to make drugs?

A

Goats

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17
Q

What are the types of biologics?

A

Vaccines
Hormones
Blood products
Cytokines
Gene and cellular therapies
Growth factors
Monoclonal antibodies
Fusion proteins

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18
Q

Where do sources of variation come from in biologics?

A

Same gene sequence
Different vector
Different expression
Different cell line
Different bioreactor conditions

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19
Q

What happens during upstream in bioreactors?

A

Sterilization
Preparation of media
Gathering raw materials

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20
Q

What happens in production of bioreactors?

A

There is free cells, immobilized cells and enzyme bioreactor

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21
Q

What happens in the downstream of bioreactors?

A

Product recovery

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22
Q

What is the most common type of bioreactor?

A

Submerged type - stir tank

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23
Q

What are the parts of a stirred-tank?

A

Sparger
Baffles

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24
Q

What are spargers used for?

A

Introducing air bubbles of optimal size to maintain homogeneity of media

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25
Q

What is baffles?

A

The metal plate used to prevent vortexing and ensure homogeneity of the culture media

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26
Q

Explain an airlift bioreactor?

A

There is an inner and outer chamber.
The inner chamber is the gassed region and the cells are lifted with air.

27
Q

Pros of a micro-carrier.

A

Can provide extremely high productivity within a compact size
Has been used widely for culture of immobilized mammalian cells
Use porous glass beads to provide a large surface area of cells

28
Q

What are the modes of bioreactor operations?

A

Batch
Continuous
Fed-batch

29
Q

Which bioreactor operation does not have an inlet or outlet component?

A

Batch operation

30
Q

What is there in bacterial proteins?

A

Inclusion bodies

31
Q

Why are inclusion bodies good?

A

Can easily be recovered to yield proteins
More resistant to proteolysis

32
Q

What will inclusion bodies dissolve in?

A

SDS
Urea
Guanidine hydrochloride

33
Q

How do you prevent proteases?

A

Add protease inhibitors

34
Q

What is glycosylation?

A

Post translational modification - attachment of a sugar molecule (oligosaccharide)

35
Q

What can glycosylation affect?

A

Pharmacological function
Immunogenicity
Solubility
Stability
Serum half life

36
Q

What are potential contaminants?

A

Host
Product
Process

37
Q

In ion exchange chromatography, if your protein has a net + charge, what kind of chromatography would you use?

A

Cation exchange chromatography

38
Q

What is the charge on a raisin in cation exchange chromatography?

A

Negative

39
Q

What will treating an extracted protein with an SDS do?

A

Put a negative charge all over the protein and unfold the protein

40
Q

What are the two ways to detect a protein?

A

ELISA or Immunofluorescent

41
Q

The primary antibody binds to the _____

A

Antigen

42
Q

The secondary antibody binds to the _____

A

Primary antibody

43
Q

Which antibody has the fluorescent in indirect detection?

A

Secondary antibody

44
Q

Which antibody has the fluorescent in direct detection?

A

Primary antibody

45
Q

What is the difference between ELISA and immunofluorescent?

A

ELISA uses and enzyme for detection and immunofluorescent uses fluorescents for detection

46
Q

What must stay the same in biosimilars?

A

Presentation
Dose
Administration mode

47
Q

What do biosimilars involve?

A

Reverse engineering

48
Q

What are the product related substances and impurities?

A

Primary structure
Biological function
Higher order structure

49
Q

What are the process related impurities?

A

Stability
Receptor binding and immuno-chemical properties
General properties and excipients

50
Q

Where is the effort for each; stand-alone product and biosimilar?

A

Stand-alone - goal is to determine clinical effect so in clinical trials
Biosimilar - goal is to determine similarity so in functional and physiochemical characterization

51
Q

Where are sources of variability for biosimilars in cloning, protein expression and production?

A

Vector and gene sequence you use
Which host you use
Cell expansion - different cell line
Different bioreactor conditions

52
Q

Where are sources of variability for biosimilars in protein purification and formulation?

A

Different operating conditions
Kind of chromatography
Which filter paper supplier
Methods of characterization and stability
Formulation

53
Q

Differences between biosimilars and generic drugs?

A

Biosimilars:
Complex
Almost impossible to fully characterize; similar but not identical to reference
Immunogenic
$100-200 million/molecule

54
Q

What are the barriers to protein drug delivery?

A

Blood brain barrier
Intestinal epithelial barrier
Capillary endothelial barrier
Enzymatic barrier

55
Q

What is passive targeting?

A

The ‘natural’ disposition pattern of the carrier system is utilized for delivery

56
Q

What is active targeting?

A

The concept where attempts are made to change the device by using “homing principle” to select one particular tissue or cell type

57
Q

When to target drugs?

A

Drugs with high total clearance
Increase in rate of elimination of free drug
Response site with relatively small blood flow

58
Q

What does the fate of particulate carriers depend on?

A

Size
Charge
Surface hydrophobicity
Presence of homing device on their surfaces

59
Q

Where should the target site be when using liposomes?

A

In the blood

60
Q

What is PEGylation?

A

Wont get taken up by macrophages - preventing immune response
The drug will have greater solubility in water
Very easy to attach targeting molecules
Decrease accessibility for proteolytic enzymes and antibodies

61
Q

What can antibiotics be produced by?

A

Bacteria, viruses, synthetic, and actinomycetes (type of bacteria)

62
Q

What is the difference between polyclonal and monoclonal antibodies?

A

Polyclonal:
Cheap to produce
Mixed populations of antibodies
May bind to different areas on target molecule
Tolerant or small changes in protein structure

63
Q

What makes something a drug candidate for MRDFs?

A

Drugs with neither high or low water solubility(in the middle would be okay)
High partition coefficient
Must be stable inside the body for extended period of time
Absorption cant be too slow or too fast
Can’t have high level of protein binding in blood(complex)
Can’t have narrow therapeutic window

64
Q

If someone is on an extended release OD tablet but the pharmacy is out of stock, should you give them a SR BID tablet or an IR TID tablet?

A

The SR BID tablet because it is the most similar to their current formulation.

PHAR 124 (4 decks)

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