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PHAR 124 > Pharmaceutical Biotechnology > Flashcards

Pharmaceutical Biotechnology Flashcards

1
Q

What are the methods of transformation that we discussed? (5)

A

Heat Shock
CaCl2 Transformation
Lipofection
Electroporation
Microinjection

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2
Q

How many elements does a plasmid vector usually contain

A

3
OriC
Selectable marker
Restriction site

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3
Q

What does rifamycin do?

A

Both inhibit transcription by binding prokaryotic but not eukaryotic RNA polymerases

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4
Q

What is a restriction in using bacterial expression systems?

A

Incapable of producing glycoproteins

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5
Q

What are primary cell lines?

A

When cells are surgically removed from an organism and placed into a suitable culture environment they will attach, divide and grow.

Shorter passage life

Not used for longterm experiments

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6
Q

What is a cell line?

A

If the cells in a cell strain undergo a transformation process (spontaneous or induced changes in growth properties) that makes them “immortal”

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7
Q

What are the common culture surfaces used for cell culturing?

A

serum,
collagen,
laminin,
gelatin,
poly-L-lysine

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8
Q

What are CHO cell lines?

A

Type of cell line that has been used for stronger expression units and has high productivity

Can grow serum free

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9
Q

What is Pronuclear microinjection?

A
  • Introduces the transgene DNA at the earliest possible stage of development of the zygote
  • DNA is injected directly into nucleus of egg or sperm
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10
Q

What is Attryn?

A

Anti-blood clotting protein which was produced from the mammary glands of transgenic goats

First drug made from genetically engineered animals approved by the FDA

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11
Q

What is Erythropoietin?

A

Treat anemia and increases RBC count

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12
Q

What is the process of protein purification and formulation?

A

Protein recovery
Purification via chromatography
Purified bulk
Protein characterization
Formulization

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13
Q

What are the three cultivaiton systems for culturing cells?

A

Free in suspension
Attached to microspheres
Immobilized states as a monolayer

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14
Q

Bioreactors may be?

A

Liquid or solid state

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15
Q

What is the role of an impeller in a bioreactor?

A

Stir the cells

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16
Q

What is a micro-carrier bioreactor?

A

Uses glass beads for cells to grow on

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17
Q

What are spargers used for in bioreactors?

A

Introduce air into stirred tank bioreactor

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18
Q

What is the most commonly used bioreactor?

A

Stirred bioreactor

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19
Q

What is the important function of baffles in a bioreactor?

A

Prevent vortexing

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20
Q

How often is a “batch” media/product harvest/culture cells changed?

A

1/1/1

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21
Q

How often is a Fed-Batch media/product harvest/culture -cells changed?

A

> 1/1/1

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22
Q

How often is a continuous media/product harvest/culture -cells changed?

A

> 1/>1/1

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23
Q

What are the advantages of inclusion bodies?

A

Resistant to proteolysis and are protected from cleavage

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24
Q

What are the disadvantages of inclusion bodies?

A

Protein bound in inclusion bodies are biologically inactive

Usually denature in SDS, Urea, Guanine Hydrochloride

Addition of mercaptoethanol or DTT will enhance the solubilization of these inclusion bodies

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25
Q

What is the steps in inclusion body breakdown?

A

Intracellular product, breakdown the cells and removal of cell debris

Extracellular product, recovery of cell culture medium by removal of cells using centrifugation or filtration

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26
Q

Proteins become much more ___ at high temperatures

A

susceptible

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27
Q

What can affect a proteins characteristic and function?

A

Solubility
Stability
Serum half life
Pharmacological function
Immunogenicity

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28
Q

Which cell culture media contains additional proteins?

A

FSC
Growth factors
hormones

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29
Q

What are the potential contaminants in serum?

A

Proteins
viruses
bacteria
prions
endotoxin
proteases

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30
Q

What most frequent source of virus introduction is?

A

animal serum

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31
Q

What are pyrogens?

A

endotoxins that are potentially hazardous substances and cannot be removed via simple sterile filtration

32
Q

What is the most common method of achieving pure protein samples?

A

Chromatography

33
Q

What are the phases of chromatography?

A

Stationary phase
Mobile Phase

34
Q

What is stationary phase of chromatography?

A

Insoluble matrix with which components of the mixture interact with during separation

35
Q

What is mobile phase of chromatography

A

Solution just moves through the column

36
Q

How many times are proteins run through a chromatography column?

A

2-3 times

37
Q

How does ion exchange chromatography work?

A

Basically where the substrate is tagged with an anion or cation

38
Q

What are the components of parenteral formulations?
SABAPA

A

Solubility enhancers
Anti-adsorption and anti-aggregation agents
Buffer components
Anti-Oxidants
Preservatives
Active ingredients

39
Q

What is often added to help insulin not be adsorbed through the skin

A

Albumin will cover the hydrophobic regions allowing a competition between albumin and insulin for the surfaces

40
Q

What are commonly used antimicrobial agents that are preservatives?

A

Thimersol, phenylmercuric nitrate, alcohol

41
Q

What is a biosimilar?

A

Copy of a commercially available biopharmaceutical

42
Q

Biosimilarity for antibodies can only be established how?

A

Through active evaluation in clinical trials and experiments with the reference product

43
Q

What are interchangeables?

A

Interchangeables can be prescribed without the interference of a health care professional

44
Q

What is the average time to produce a biosimilar?

A

7-8 years compared to 8-10 years

45
Q

What are some barriers to protein drug delivery?

A

Enzymes
Intestinal barrier
Capillary barrier
Blood brain barrier

46
Q

How can we increase the permeability of the absorption barrier?

A

Additon of FA/Phospholipids

liposome delivery

Decrease peptidase activity at the site of absorption

47
Q

What is iontophoresis?

A

electric pads on skin that allows the drug to follow from positive to negative. Gets caught in blood stream and moves

Pulsatile secretion

48
Q

What are the types of targeting systems we can use

A

Passive targeting (Macrophages)
Active targeting

49
Q

What is PEGlyation?

A

Prevents an immune response and increases the ability of a drug to be soluble in water?

50
Q

What is a disadvantage of liposomes?

A

Poor access to targets outside of blood
High resistance

51
Q

What are the controlled released systems for parenteral delivery?

A

open loop and closed loop

Continuous infusion pump vs biosensory pump combo

52
Q

What is an antisense oligionucleotide?

A

Short DNA analogue that hybridizes with the complementary mRNA

53
Q

What does hybridization lean to?

A

Inhibition of gene expression

54
Q

What is Luxturna?

A

First DFA approved gene therapy for retinol dystrophy Very expensive

55
Q

What are hybridomas?

A

These are cancer cells fused with antibody-forming cells to produce more antibodies

56
Q

Which phase are primary metabolites produced?

A

Log phase

57
Q

Which phase are secondary metabolites produced?

A

Late log phase and early stationary phase

58
Q

What is considered low risk in the manufacturing process?

A

Change filter supplier, Move equipment within the same facility

59
Q

What is considered moderate risk in the manufacturing process?

A

Move to new production facility

60
Q

What is considered high risk in the manufacturing process?

A

Change cell culture media, New cell line or major formulation change

61
Q

Biosimilars are not

A

Second generation or generic drugs

62
Q

What is the MOA of rifamycin?

A

Inhibit chain elongation of the RNA strand.

63
Q

What are bacteria not capable of producing?

A

glycoproteins

64
Q

What is Hayflick’s phenomenon?

A

Describes how primary culture cells have a limited number of passages before the cells just simply stop producing

65
Q

What year did the FDA approve atttryn?

A

2009

66
Q

What are the three parts of a biological process? with respect to bioreactors

A

Production, Product recovery, Sterilization of raw materials/air

67
Q

Most bioreactors used in industry are?

A

Submerged type or liquid

68
Q

The function of the impeller?

A

Stir it up

69
Q

The function of the baffle?

A

Get rid of em vortexes

70
Q

Function of the Sparger

A

Bubbles bubbles

71
Q

What are micro-carrier bioreactors commonly used for?

A

Culture of immobilized mammalian cels

72
Q

What are the reducing agents to break disulfide bonds?

A

Mercaptoethanol
Dithiothreitol

73
Q

How can solubility be improved for protein delivery?

A

Proper PH
Add Lysine or Arginine
Surfactants
Sugars and glucose

74
Q

Albumin is both an anti

A

Adhesion and anti aggregation

75
Q

What is a limitation of osmotically driven systems/

A

Fixed release rates which isnt always desired

76
Q

Antisense oligionucleotides can affect

A

Inhibition of 5’ cap formation,
RNA splicing
Activation of RNASE H
Block ribosomal subunit

77
Q

How expensive is therapeutic antibodies for the treatment of disease?

A

300+ billion

PHAR 124 (6 decks)

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