Many novel immunoassays methods employed functional (antibodies-conjugated) nanoparticles as novel probe to detect analytes. My question is how to determine the number of antibodies conjugated on the nanoparticles? Can anyone answer this question？
In my opinion is not important to know the number of antibodies that coat the nanoparticle surface, but the efficiency with which recognize the antigen, since some of them may be misguided in their own bonding and hidden areas of antigen binding.
You need to know the molecular weight of your antibody. Also from ICP-OES you can determine the concentration of AuNPs, subsequently you can calculate the number of AuNPs particles in given solution.
Although, to me it is not necessary to calculate the number of antibody binds to AuNPs. Instead of you can calculate the amount of antibody is conjugated with AuNPs.
Hello, i use ADECA methods which permits to estimate directly the antibody grafting.
The easiest way to calculate the amount of a loaded protein on gold particles is to measure the concentration of the protein before and after conjugation using a nanodot plate reader. The size and concentration of thee particles also can be measured using either UV vis or TEM.