Dissociation constants
Smaller Kd = stronger affinity
| Antibody/Receptor | Antigen/Ligand | Kd |
|---|---|---|
| PirB | Beta-amyloid oligomers | 180 nM |
| LilrB2 | Beta-amyloid oligomers | 206 nM |
| Gantenerumab | Beta-amyloid oligomers | 1.2 nM |
| HyHEL-10 | Chicken lysozyme | 0.67 nM |
| VRC01 | gp120 | 3.88 nM |
Gantenerumab binding to other things:
Beta-amyloid oligomers Beta-amyloid fibrils Beta-amyloid monomers | 1.2 nM 0.6 nM 17 nM |
Detecting antigen binding
We can't directly label gp120 or chicken lysozyme with biotin or fluorophores because conjugation alters lysine residues, which are critical to binding between VRC01 and gp120 and between HyHEL-10 and chicken lysozyme. This doesn't affect Gantenerumab's interaction with beta-amyloid oligomers because this interaction doesn't involve lysine residues (also, in this paper they use biotinylation to perform SPR between Gantenerumab and beta-amyloid oligomers).
Options are
- biotinylated antibodies (that we can use with the AF594-Streptavidin we already have)
- fluorophore-conjugated antibodies (preferably AF594)
- primary antibodies that we can combine with secondary antibodies we already have (see list here)
Gantenerumab/beta-amyloid: Biotinylated beta-amyloid oligomers + Alexa fluor 594-conjugated Streptavidin
VRC01/gp120:
| Name | Company | Species | Clonality | Valid for | Conjugated? | Price |
|---|---|---|---|---|---|---|
| Anti-HIV1 gp120 antibody (Biotin) (ab53937) | abcam | Goat | Polyclonal | WB, ELISA, IHC-Fr, ICC/IF | Biotin | 100uL/$378 |
HyHEL-10/chicken lysozyme:
| Name | Company | Species | Clonality | Valid for | Conjugated? | Price |
|---|---|---|---|---|---|---|
| Anti-Lysozyme antibody (ab391) | abcam | Rabbit | Polyclonal | WB, IP, ELISA, ICC | No, could use with 594 donkey anti-rabbit secondary | 100ug/$373 |
| lysozyme antibody (FITC) orb189142 | biorbyt | Rabbit | Polyclonal | IF/ICC | FITC | 100ug/$315 2-3wk wait |