Biological Agents 1

Question 1

Name the 2 types of Pharmacological agents

Answer 1

1) NCE’s - New Chemical entities

2) NBE’s - New Biological entities (aka; Biologics, Biological agents)

Question 2

Give 2 examples of small molecules drugs and their molecular weight

Answer 2

1) Aspirin - 180

2) Insulin - 5080

Question 3

Name 4 unexpected properties of Protein drugs

Answer 3

1) Aggregation (side effects)
2) Incorrect folding
3) Amino Acid (AA) modifications
4) Truncation and Proteolysis

Question 4

Name expected properties of Protein drugs

Answer 4

• Size, charge and Hydrophobicity
• Correct folding (S-S bonds)
• Glycosylation
• Bioactivity
• Subunits

Question 5

Name 6 types of biologics

Answer 5

1) Native proteins
2) Recombinant proteins
3) Vaccines
4) Peptides
5) RNAi & microRNA
6) Gene therapy/Viral vectors

Question 6

Name 2 types of recombinant proteins

Answer 6

1) Wild type proteins

2) Genetically engineered proteins

Question 7

What do gene based/cellular biologics do ?

Answer 7

Treat medical conditions no other treatments are available for

Question 8

Give a few differences between biologic and chemical agents

Answer 8

Biologics - Larger, More complex, Dilute, Made by living cells, Aqueous solvent required, Chromatography required for Isolation/Purification

Chemical - Smaller, Less complex, Concentrate, Made from multiple chemical reactions, Organic solvent required, Crystallization required for Isolation/Purification

Question 9

Name 4 ways Protein drugs differ from Small molecule drugs

Answer 9

1) they may contain Intrinsic infectious agents
2) Aseptic techniques required for production
3) Exact structure may be unknown
4) Heterogeneous (different) compositions = impurities produced

Question 10

Name the 4 processes of Biological drugs

Answer 10

1) Expression - What system ?
2) Fermentation - Complexity, Control, Cell density etc
3) Purification - Yield, Efficiency
4) Characterisation - Purity, Identity, Potency

Question 11

What are the 5 major features of an eukaryotic expression vector?

Answer 11

1) Promoter (elongation of Factor 1 alpha)
2) MCS (cloning sites)
3) AmpR (ampicillin resistance)
4) IRES (Internal Ribosome Entry site)
5) Puromycin resistance (pac)

Question 12

What does the presence of IRES sequence create ?

Answer 12

Creates a Bicistronic expression vector

Question 13

Name 3 ways modified DNA can be introduced into a desired cell

Answer 13

1) Electroporation
2) Lipid transfection
3) Calcium phosphate

Question 14

Name pros and cons of mammalian cell production ?

Answer 14

Con - Not easy and cheap to grow the cells (due to authentication and contamination issues)

Pro’s - May produce large amounts of product
- Affinity tags make Purification straight forward
- Protein = likely to have suitable PTM’s and is
produced in fully folded active form

Question 15

Name 4 examples Post translational modifications (PTMs)

Answer 15

1) Glycosylation (n linked or O linked)
2) Proteolysis - RXXR sites (R = arg)
3) Phosphorylation (on tyr, ser and thr)
4) Disulphide bond formation (covalent dimer formation)

Question 16

Name one difference between a microbial fermenter and a mammalian cell bioreactor ?

Answer 16

Animal cell bioreactor has a ‘Marine type’ impeller whereas microbial fermenter has a Rotating impeller