For my Lateral Flow Assay I'm trying to increased the concentration of the biotinylated antibody conjugated with the 40 nm Streptavidin gold nanoparticles, but with a higher concentrations of 0.1 ug/ml the particles are aggregating. Normaly I dilute my antibody in this buffer:20 mM Sodium borate (ph 8), 1%BSA, 2% Sucrose, 0.02% sodium azide.The centrifugations have the follow conditions: 9000g,6min, 4ºC. I already try dilluted the antibody in PBS+1%BSA, and in PBS in the following conditions: 2500g,20min,4ºC, for a concentration of biotinylated antibody of 0.3 ug/ml. However for this concentration the particles aggregated. Can you help me ?
DEar Maria Pereira Pereira
how did you biotinilated the antibody?
Did you use limited biotin amount to do not obtain multiple biotinilation? Since a multibiotinilated antibody may act as a bridge between different particles and induce aggregation.
best regards
Manuele
Decrease the extent of biotin loading on your antibody. How much biotin are you loading right now?
Is there a reason when you can't adsorb the MMP9 antibody to the gold particles directly? Is using biotin-avidin strictly necessary?
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts