Biologicals

Question 1

Proteins vs Small Molecules

Answer 1

See separate sheet

Question 2

How are therapeutic proteins normally made?

Answer 2

Relies largely on recombinant DNA methods (apart from small peptides)

Question 3

Humanized antibodies contain…

Answer 3

Murine complementarity determining regions

Question 4

What are the most common hosts for the production of therapeutic proteins in cost order (cheapest first)

Answer 4

Bacteria
Yeast
Mammalian Cells

Question 5

How easy is it to scape up the various hosts for the production of therapeutic proteins?

Answer 5

Bacteria & Yeast
- Relatively easy - bigger vat
Mammalian Cells
- More difficult and expensive
- Cells that grow in suspension are easier
- Many cell lines are adherent - cell factories or microcarriers can be used

Question 6

Which operations may denature or aggregate proteins

Answer 6

• Freezing/Thawing
• Agitation (interfaces)
• Sonication
• Contact with silicone oil
• Low or high pH
• Low or high salt
• Specific salts
• Heat

Question 7

Give examples of interfaces that can cause aggregation?

Answer 7

Air-water
- Vials, IV bags
Oil-water
- Silicone-coated syringes
Hydrophobic surfaces
- IV set & bag

Question 8

Is Normal Saline usually ok as an IV diluent?

Answer 8

Yes

Some hydrophobic proteins are less soluble/insoluble at high salt
Half normal may be better?

Question 9

Is Dextrose usually ok as an IV diluent?

Answer 9

No

• Can react with lysines on the protein surface to form Schiff’s bases relatively quickly at ambient temps
• Can occur in vivo, especially in patients with uncontrolled diabetes

Question 10

What is Tissue Engineering

Answer 10

Aims to replace diseased or damaged living tissue with living tissue designed and constructed for the needs of each individual