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Introduction to Biopharmaceuticals & Gene Therapy > Biopharmaceuticals > Flashcards

Biopharmaceuticals Flashcards

1
Q

What are pharmaceuticals?

A

Drugs used to diagnose, prevent, treat or cure disease

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2
Q

What is a biotechnology medicine/product?

A

Pharmaceutical product used for therapeutic or in vivo diagnostics, produced in full or in part by modern biotechnological means (include antibiotics extracted from fungi; insulin from pig pancreas or recombinant insulin)

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3
Q

Give examples of products which are from a animal origin

A

Antibodies, enzymes, hormones, blood factors, growth factors, cytokines

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4
Q

Give examples of products which are form a plant origin

A

aspirin, alkaloids, digitalis, quinine

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5
Q

Give example of products form a microbial origin

A

vaccines, antibiotics, toxins/toxoids

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6
Q

What are biopharmaceutical proteins or nucleic acids?

A

based pharmaceuticals that are used for therapeutic or in vivo diagnostics, produced by means other than direct extraction from native biological source

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7
Q

Give examples of pharmaceuticals from chemical synthesis?

A

Paracetamol (acetaminophen), ketamine, acyclovir

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8
Q

Give examples of pharmaceuticals from biological extraction

A

Factor VIII, insulin, alkaloids (morphine), antibiotics, vaccines, antibodies (passive immunity), thrombolytic agents

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9
Q

Give examples of some biopharmaceuticals

A

Factor VIII, insulin, vaccines, antibodies (e.g. monoclonal), thrombolytic agents

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10
Q

What are the advantages of biopharmaceuticals?

A

Overcomes issue of source availability (e.g. Cytokines)

Improves product safety (e.g. Blood products – HIV, hepatitis, CJD)

Alternative to extraction (e.g. Insulin from slaughterhouses, hormones from urine)

Engineering of more effective therapeutic proteins (e.g. Insulin, Factor VIII)

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11
Q

What region of a chromosome is the encoding region?

A

euchromatic gene

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12
Q

What fraction of the genome is transcribed into RNA?

A

1/3

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13
Q

What % of RNA is believed to produce proteins?

A

~5%

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14
Q

What are bioinformatics used for?

A

to identify functional sequence motifs (e.g. catalytic sites, transmembrane regions) by homology to other known genes (homologues), even in different species (orthologues)

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15
Q

What are knock-out animals?

A

generation of animals in which the study gene is deleted – observation of phenotype can Often reveal role of gene product

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16
Q

What are the different variations in gene expression between cells?

A

different cell types - stage in organisms development - phase of cell cycle - response to internal and external (i.e. environmental) factors

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17
Q

What are gene chips?

A

study of differential RNA expression (transcriptome) in different cells/conditions (e.g. Expression in cancer cell may provide drug target)

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18
Q

What are proteomics?

A

Proteomics: analysis of cellular protein expression (proteome)

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19
Q

What are structural genomics?

A

3D protein analysis (NMR) – structure may predict function

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20
Q

What is pharmacogenomics?

A

correlation of gene sequences to drug response

Determine which drugs (& dose) to give which individuals – benefits v side effects

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21
Q

What do pharmacogenomics look at?

A

mutations and polymorhisms (single nucleotide polymorphisms – SNPs)

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22
Q

what are most BPs produced by?

A

genetic engineering using recombinant expression systems

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23
Q

What are the steps of using recombinant expression systems?

A

First - identification and isolation of therapeutic gene/custom synthesis

Molecular cloning into vectors (e.g. plasmids, viruses)

Propagation (maybe protein expression) in cell lines

Purifying products (DNA, protein, vector)

24
Q

What is used to clone mRNA into cDNA?

A

produced using retrovirus reverse transcriptase

25
Q

What regulates the expression of genes?

A

Gene promoters

26
Q

What does differential splicing allow?

A

Different mRNAs and thus proteins to be formed from the same gene sequence

27
Q

What are introns?

A

Non coding DNA

28
Q

What is the gene often fused to in order to ease purification?

A

A leader sequence or tag that allows the protein to be isolated using, for example antibodies

29
Q

What are the two types of expression a cell line can have?

A

Transient/stable expression

30
Q

What cell lines are most BP proteins produced in?

A

E.coli (e.g. K12) or mammalian cell lines (e.g. Chinese hamster ovary, CHO; baby hamster kidney, BHK)

31
Q

What does poor post-translational modifications affect?

A

Functions

32
Q

What is removed form microbial cell lines?

A

Surface LPS pyrogenic

33
Q

What can high-level intracellular expression of heterologous proteins produce?

A

Inclusion bodies

34
Q

What is a advantage of inclusion bodies?

A

Can aid centrifugal privation

35
Q

What is a disadvantage of inclusion bodies?

A

Aggregates of partially folded foreign proteins

36
Q

What is a solution of inclusion bodies?

A

express soluble intracellular fusion protein/tag (foreign-native protein)

Purify and enzyme cleave off target protein

37
Q

What are some examples of fusion proteins?

A

Fusion proteins include glutathione transferase, thioredoxin, maltose-binding protein

38
Q

What are advantages of culturing yeast cell line?

A

Cultured relatively fast and cheaply

39
Q

What is the most widely used yeast cell line?

A

S. cerevisiae most widely used (e.g. insulin, Hepatitis B vaccine antigen, anticoagulants)

40
Q

What are advantages of using S. cerevisiae?

A

Genetics well characterised

Safe – used in brewing, baking

41
Q

What does Mannose glycosylation lead to?

A

rapid blood clearance

42
Q

What are disadvantages of mammalian cell lines?

A

Slower and more expensive to set-up, up-scale and maintain (nutrients, fragility)

43
Q

What are most widely used mammalian cell lines?

A

Chinese Hamster Ovary (CHO), Baby Hamster Kidney (BHK) most widely used (e.g. insulin, vaccines). Hybridoma cells for monoclonal antibodies

44
Q

How are transgenic animals produced?

A

by microinjecting exogenous DNA construct into egg cells

45
Q

What are some advantages of transgenic animals?

A

Milk safe, well characterised, abundant

Continued inheritance of recombinant DNA in every cell

Relatively low costs after initial transgenic outlay

46
Q

What is a disadvantage of transgenic animals?

A

significant lag period from viable embryo and manufacture (n.b retrovirus vectors)

47
Q

What is the only licensed transgenic animal BP?

A

human antithrombin ‘Atryn’

48
Q

Give a example of some glycosylation differences using transgenic animals?

A

c.f. CHO cells – seemed to enhance efficacy but reduce half-life

49
Q

What do Agrobacteria species contain?

A

Ti plasmids

50
Q

What are advantages of transgenic plants?

A

Cultivation/harvest/scale up easy and costs low

Proteins usually stable

Plants free of human pathogens

Can get low expression/gene silencing

51
Q

What are disadvantages of transgenic plants?

A

Glycosylation significantly different – immunogenic

Potential plant metabolite contamination (alkaloids)

Public issues surrounding GM crops – cell lines?

52
Q

What is CaroRX?

A

recombinant ‘plantibody’ against Strepococcus mutans in tooth decay - topical BP

53
Q

What is Merispace?

A

oral (corn) recombinant gastric lipase to treat lipid malabsorption due to poor pancreas secretion (e.g. CF)

54
Q

What can Agrobacteria and plant viruses also be used for?

A

Transferring BP genes into plants transiently

55
Q

What are advantages of insect cells?

A

High level expression (strong promoters) – up to 50% of cell protein

Rapid and cheaper culture than mammalian cells

Most human pathogens don’t infect insect cells

56
Q

What are disadvantages of insect cells?

A

Most protein is intracellular – purification needed

Often incomplete/markedly different glycosylation

57
Q

Cervarix HPV cervical cancer vaccine particles from?

A

recombinant L1 capsid protein

Introduction to Biopharmaceuticals & Gene Therapy (4 decks)

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