AB / Recombinant Protein Development (Uppalapati) Flashcards
The first MAB was created in what year?
1975
The pharmaceutical market share predicts that antibody drugs will be valued at how many billions of dollars by 2025?
$300bil
The AA sequence determines what level of protein structure?
Primary
Secondary
Tertiary
Quaternary
Primary
Alpha Helices and Beta Sheets are examples of what level of protein structure?
Secondary
Protein complexes are an example of what level of protein structure?
Quaternary
3D structures of proteins are an example of what level of protein structure?
Tertiary
What immune cell is considered to be the antibody factory?
B Cells
What portion of the antibody elicits effector functions once it binds to target receptors?
Fc
What holds the hinge region of an antibody together?
Two Disulfide Bonds
What are the effector functions of antibodies?
-Neutralize pathogens
-Link antigens together
-Opsonization (ie. Lysosome uptake & digestion)
-Activate compliments
-AB Dependent, Cell-Mediated Cytotoxicity
How can antibodies be employed in Radioimmunotherapies?
As a targeting vector (deliver radioactive drug to target site in the form of an AB-Drug Conjugate).
How does Bevacizumab work?
IgG AB that binds to VEGF & prevents this growth factor from crossing epithelial membranes (stops Angiogenesis).
How do Immune Checkpoint Inhibitors such as Ipilimumab (CTLA4) & Nivolumab (PD1) work?
Bind & block respective receptors, rendering them unable to bind to targeted ligands… This reverses immunosuppression & allows T Cells to elicit immune response!
CAR-T & BITE therapies serve to treat cancers how?
Recruitment-based therapies that trigger / ramp up immune response.
What are the two most important factors that influence protein-protein binding?
1) Are the two proteins structurally compatible?
2) Are AAs of the two proteins complementary to each other?
Most AB variation occurs within _______ ______ ______.
Hint: This segment of the AB also mediates antigen binding.
Complementary Domain Regions
If we neglect junctional flexibility & nucleotide additions or deletions, how many different combinations of heavy / light chains are possible?
2.64 x 10^6
What bacteriophage is dubbed the “workhorse for protein engineering”?
M13
What disadvantage does a Hybridoma Mouse have over a Xenomouse?
Mouse AB locus is expressed (generates immunogenic response in humans)… In Xenomouse, the mouse AB locus is inactivated (so no immunogenic response to therapy).
How does a Canonical AB differ from an AB-Drug Conjugate?
Canonical: AB alone is the drug!
AB-Drug Conjugate: AB carries drug to target site.
What advantage does the small size of AB Fragments have in terms of drug targeting / delivery?
Enables deeper tumor penetration!
Are the success rates of clinical trials for small molecule drugs greater than MABs?
Nope… MABs = 23% (versus 7% for small molecule drugs).
What cancer signaling receptor / ligand combination saw the greatest amount of MABs developed for it?
PD1 / PDL1
Aside from cancer indications, what else can Bevacizumab be used for?
Age-Related Macular Degeneration
What types of roles do BITE ABs possess?
-Bridge (proximity)
-Prevent Rec Dimerization
-Activate Rec
-Piggybacking
Which of the following recombinant AB technologies is oldest? Newest?
Knob-in-holes
Quadroma
Cross-Mab
Bispecific
Oldest: Quadroma
Newest: Bispecific
Advantage that Cross-Mab technologies have over Knob-in-holes?
Prevents LC Crossover (as well as HC Crossover)… Knob-in-holes only prevents HC Crossover.
Which Bispecific AB therapy has proven to be most successful? What does it treat?
Emicizumab; Hemophilia
Do ABs or Alternative Scaffolds have better tissue penetration?
Alt Scaffolds
Do ABs or Alternative Scaffolds demonstrate enhanced stability?
ABs… Alt Scaffolds have poor stability.
Which is cheaper to produce: ABs or Alt Scaffolds?
Alt Scaffolds… ABs very expensive.
Which demonstrates immunogenicity: ABs or Alt Scaffolds?
Alt Scaffolds
