Dear colleagues involved in the subject of the use of for coupling antibodies onto surface of the gold nanoparticles, please, provide me the information of the biblical studies known to them, from which I could establish, how the number of antibodies coupled onto surface of the gold nanoparticle have influence to the sensitivity of the LFIA method.
Article How Antibody Surface Coverage on Nanoparticles Determines th...
Dear Anna, there is no easy answer to this as there are plenty of confounding factors, as e.g. polyclonal vs. monoclonal antibodies, coating (=passive absorption) vs. chemical coupling, affinity, valency of your analyte, particle size, ...
Generally, the more surface coverage the better. However, there are cases where a single antibody per gold particle also works well. Have you tested the performance of your antibody in other assays? How well do you know the antibody you are working with? And what is the purpose of your experiment, i.e. which Analyte do you want to detect?
Dear Mr. Sack, I work with competative LFIA, with monoclonal Ab, I have tested the Ab by Indirect ELISA. Particles size 40 nm. Sensitivity from ELISA is good. I did covalent coupling and also passive absorbtion, for passive absorption we need more Antibody, I was wondering, should I expect lease sensitivity because of amount of Antibody.
Dear Anna, you need to test the differences, it is impossible to predict the performance of the antibody after coupling as every antibody is unique. Some antibodies get inactivated during coupling as their binding site gets blocked, others work just perfect. So you need to test this (each time).
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