I'm developing a lateral flow assay at present and I've had trouble with my antibody-gold conjugate not moving off my conjugate pad. When I run test solutions through it (PBS, PBS + 1% Tween20, PBS + 1% BSA) the nanoparticles remain on the conjugate pad and do not wick up the nitrocellulose with the rest of the solution.
Conjugate pad is made of glass fibre - GFDX from Merck Millipore. Antibody was conjugated with Cytodiagnostics 40nm kit purchased from Sigma Aldrich. I've tried directly spotting the antibody complex onto the glass fibre or pretreating it with PBS + 1% Tween20 + 1% BSA and then spotting it on. I don't have an aerosol sprayer setup in my lab unfortunately.
Thanks in advance for any help!
How did you dry the gold conjugate on the pad? I do not see any mention of sucrose. It is very important that you have sucrose or trehalose in the gold conjugate drying buffer at 1-5%. Sucrose aids in the re-hydration of the dried gold-conjugate. Also, I believe that you are using too much Tween-20 at 1%. Again in my experience, This detergent does not re-hydrate easily and I would go for Tween-80
Add some detergent in Glass fiber pad treatment buffer like Brij, triton x100. Sucrose use for stability it no role with conjugate movement. Reduce tween 20. Check you conjugate. If it aggregated then not possible to move. If need more information please call me on my whatapp number +917807279791
Triton x100 and high concentration of glucoe and sucrose should help
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