Best way to determine if gold nanoparticles will bind with HE4 antibody?
Anyone have helpful protocols?
Gold particles will bind most antibodies and most people just use the gold conjugate in the final application to see if the binding was successful. You can do passive binding, in which case you will need to try different pH conditions, but usually above pH 8 will be best. Or you can do covalent binding which is generally far easier. What size of gold particle do you need, as this will affect how you go about this task?
I guess I am not sure about the size I will need. I plan on binding the gold particle to a nitrocellulose substrate as well too.
What would you recommend? Do you have good protocols to go by?
Thank you for your response!
If you need something in the range 10-80nm then this is probably your easiest option:
https://www.expedeon.com/products/immunoreagents/particles-particle-conjugation-kits/innovacoat/particle-conjugation-kits-innovacoat/
To prove you have antibody on the gold you can use a simple lateral flow test:
https://www.expedeon.com/products/immunoreagents/checkgo/conjugate-checkgo/
If you want a different size you will have to buy the appropriate colloid and do passive binding. This is far more demanding than covalent kits.
I think that size range should work. I am working on using paper microfludics if possible, but this is my schematic idea so far. I am not sure how feasible it all is.
1) Not sure gold nanoparticles are bound to the nitrocellulose?
2) Not sure if I need 2 antibodies for HE4 ordered in a specific way
Thank you for your links and response!
Hi Emily.
You can check the binding of antibody with gold nanoparticles by using UV-Vis spectrophotometer.The gold nanoparticles exhibited a characteristic surface plasmon resonance peak at 520 nm. which exhibited a red shift upon addition of antibodies. This can also be confiremd by gel retardation assay. Furtehr, the protein corona due to antibody can also be observed in Transmission electron microscopy.
Best
Hi Emily,
I do agree with Prabir. I have used GNP for immunolabelling with a certain antibody. The reason was to detect the antigen localization using TEM " Transmission electron microscopy".
Good luck
Emily Kight.
If you want to produce Lateral flow test, We can develop test together. I had developed various Rapid during last 10 year. If you interested email me at [email protected]
See: https://www.researchgate.net/post/what_factors_must_be_considered_in_nano_particles-protein_conjugation
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