Is there a procedure or mathematical formula to calculate the number of antibody molecules bound to a gold nanoparticle? I used PEG-EDC-NHS linking chemistry to bind antibody to nanoparticle.
You can consider measuring the unbound antibody by separating the gold nanoparticle after linking procedure followed by spectrophotometric protein measurement of the supernant to infer the amount of particle-bound antibody.
I have several suggestions:
1.you can try BCA assay to quantify the total proteins on your NPs. Use unconjugated gold nanoparticles as control.
2. Use detergent to rinse off the antibodies and perform ELISA /BCA/WB.
3. Fluorescently tag your antibodies using organic dyes, which should be fast and relatively easy to do, then measure the fluorescence of your NPs, again use unconjugated NPs as control.Add your answer
Hope it helps.
Better measure unbound antibody concentration by protein estimation and calculate from total amount added.
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