For the LFA, detection antibodies need to bound to gold nanoparticles. what are the analytical techniques to verify a successful conjugation? Please suggest any articles with the standard protocol.
Assuming there were no other protein, you can do BCA on the nps and on the reaction volume to ensure that the antibody is present at a known concentration on the beads.
Lora Benoit thank you very much for your suggestion. in BCA assay does the Nanoparticles interfere with the assay chemically/electromagnetically?
There should be a subtle change in the colour when antibodies bind. It’s not always easy to see by eye, but you can do a wavelength scan before and after addition of the antibody. There will be a slight rightward shift of the peak situated at about 524nm.
Another standard test is to stress the conjugate by adding salt. Naked particles or poorly conjugated particles will aggregate and turn blue, grey or black.
Nick Gee thank you for your valuable inputs, can we use the salt stress test for both covalent conjugation as well as passive conjugation?
Yes for passive conjugation. This is really what the test is for.
It is harder to say for covalent conjugation as sometimes the particles will already have some sort of coating layer to which functional groups have been attached. You can always try the salt stress test prior to covalent conjugation to see what happens.
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