AB / Recombinant Protein Development (Uppalapati)

Question 1

The first MAB was created in what year?

Answer 1

1975

Question 2

The pharmaceutical market share predicts that antibody drugs will be valued at how many billions of dollars by 2025?

Answer 2

$300bil

Question 3

The AA sequence determines what level of protein structure?

Primary
Secondary
Tertiary
Quaternary

Answer 3

Primary

Question 4

Alpha Helices and Beta Sheets are examples of what level of protein structure?

Answer 4

Secondary

Question 5

Protein complexes are an example of what level of protein structure?

Answer 5

Quaternary

Question 6

3D structures of proteins are an example of what level of protein structure?

Answer 6

Tertiary

Question 7

What immune cell is considered to be the antibody factory?

Answer 7

B Cells

Question 8

What portion of the antibody elicits effector functions once it binds to target receptors?

Answer 8

Fc

Question 9

What holds the hinge region of an antibody together?

Answer 9

Two Disulfide Bonds

Question 10

What are the effector functions of antibodies?

Answer 10

-Neutralize pathogens

-Link antigens together

-Opsonization (ie. Lysosome uptake & digestion)

-Activate compliments

-AB Dependent, Cell-Mediated Cytotoxicity

Question 11

How can antibodies be employed in Radioimmunotherapies?

Answer 11

As a targeting vector (deliver radioactive drug to target site in the form of an AB-Drug Conjugate).

Question 12

How does Bevacizumab work?

Answer 12

IgG AB that binds to VEGF & prevents this growth factor from crossing epithelial membranes (stops Angiogenesis).

Question 13

How do Immune Checkpoint Inhibitors such as Ipilimumab (CTLA4) & Nivolumab (PD1) work?

Answer 13

Bind & block respective receptors, rendering them unable to bind to targeted ligands… This reverses immunosuppression & allows T Cells to elicit immune response!

Question 14

CAR-T & BITE therapies serve to treat cancers how?

Answer 14

Recruitment-based therapies that trigger / ramp up immune response.

Question 15

What are the two most important factors that influence protein-protein binding?

Answer 15

1) Are the two proteins structurally compatible?

2) Are AAs of the two proteins complementary to each other?

Question 16

Most AB variation occurs within _______ ______ ______.

Hint: This segment of the AB also mediates antigen binding.

Answer 16

Complementary Domain Regions

Question 17

If we neglect junctional flexibility & nucleotide additions or deletions, how many different combinations of heavy / light chains are possible?

Answer 17

2.64 x 10^6

Question 18

What bacteriophage is dubbed the “workhorse for protein engineering”?

Answer 18

M13

Question 19

What disadvantage does a Hybridoma Mouse have over a Xenomouse?

Answer 19

Mouse AB locus is expressed (generates immunogenic response in humans)… In Xenomouse, the mouse AB locus is inactivated (so no immunogenic response to therapy).

Question 20

How does a Canonical AB differ from an AB-Drug Conjugate?

Answer 20

Canonical: AB alone is the drug!

AB-Drug Conjugate: AB carries drug to target site.

Question 21

What advantage does the small size of AB Fragments have in terms of drug targeting / delivery?

Answer 21

Enables deeper tumor penetration!

Question 22

Are the success rates of clinical trials for small molecule drugs greater than MABs?

Answer 22

Nope… MABs = 23% (versus 7% for small molecule drugs).

Question 23

What cancer signaling receptor / ligand combination saw the greatest amount of MABs developed for it?

Answer 23

PD1 / PDL1

Question 24

Aside from cancer indications, what else can Bevacizumab be used for?

Answer 24

Age-Related Macular Degeneration

Question 25

What types of roles do BITE ABs possess?

Answer 25

-Bridge (proximity)
-Prevent Rec Dimerization
-Activate Rec
-Piggybacking

Question 26

Which of the following recombinant AB technologies is oldest? Newest?

Knob-in-holes
Quadroma
Cross-Mab
Bispecific

Answer 26

Oldest: Quadroma
Newest: Bispecific

Question 27

Advantage that Cross-Mab technologies have over Knob-in-holes?

Answer 27

Prevents LC Crossover (as well as HC Crossover)… Knob-in-holes only prevents HC Crossover.

Question 28

Which Bispecific AB therapy has proven to be most successful? What does it treat?

Answer 28

Emicizumab; Hemophilia

Question 29

Do ABs or Alternative Scaffolds have better tissue penetration?

Answer 29

Alt Scaffolds

Question 30

Do ABs or Alternative Scaffolds demonstrate enhanced stability?

Answer 30

ABs… Alt Scaffolds have poor stability.

Question 31

Which is cheaper to produce: ABs or Alt Scaffolds?

Answer 31

Alt Scaffolds… ABs very expensive.

Question 32

Which demonstrates immunogenicity: ABs or Alt Scaffolds?

Answer 32

Alt Scaffolds