Exam III: Trojan Horse Paper Flashcards
What is the genome of Ebolavirus? What family is it a part of?
neg ssRNA
enveloped
Filoviridae family
What are the 5 different species of ebolavirus which have been isolated?
- Sudan
- Zaire
- Tai Forest (Ivory Coast)
- Reston
- Bundibugyo
What is the mechanism of attachment/penetration/uncoating of EboV?
EBV has glycoprotein on it envelope which binds to receptor > receptor-mediated endocytosis > GP is cleaved > NPC1 binds to GPcl > viral and plasma membrane fusion > genome release into cytoplasm
What is ZMapp?
It is the current treatment for EBV
3 humanized antibodies that target the 3 Ebola glycoproteins
What are the two locations you could add an extra VH/VL? How?
To the end of the variable regions: CJDVv
To the end of the constant region: CvJDV
What was the hypothesis of the authors of the Trojan Horse paper?
A bispecific antibody will be able to bind to EBV, be taken into the endosome, bind to NPC1, and inhibit release of the viral genome
What 2 bispecific antibodies did they create?
- FVM09~548 (engages GP and NPC1)
2. FVM09~MR72 (engages GP and GPcl)
How do you test binding affinity of an antibody to an antigen?
The higher the affinity of the protein for the target, the lower the concentration required the higher the affinity
How do you test antibody neutralizing activity?
Plot %Viral infectivity as a function of antibody concentration
How do you test the movement of antibodies (ie into the endosome)?
tag the virus in green
tag the antibody in pHrodo Red = only floureces red at low pH
green + red = yellow = colocalization
How do you test antibody effectivity as a treatment in vivo?
Create kill curves
% survival as a function of days post-infection treated with different antibodies
Why did FVM~548 not work in mice?
NPC1 is human-specific
