Looking to make gold nanoparticles and wondering which is the best way to yield enough to make several LFA tests.
Is the Turkevich method best?
I made a bear bones protocol attached. Not sure about the dilution before conjugation step. Also not sure at what temp to dry the membrane.
Hi Emily,
I believe that the simplest but still effective methodology for synthesizing AuNPs with a diameter between 10 and 25 nm is by Turkevich-Frens. For larger diameters, seeded growth synthesis may be a good option.
Since AuNPs are stabilized by citrate, if you conjugate with some molecule for the LFA, it may be interesting to use Tween 20 to prevent the nanoparticles from aggregating. For dilution, maybe you can do it serially to test the combination and the final coloring of the solution.
I hope it helped you.
You can refer to the references to prepare gold NPs with a size ranged from 3-13 nm.
1. DNA-controlled dynamic colloidal nanoparticle systems for mediating cellular interaction
2. DNA assembly of nanoparticle superstructures for controlled biological delivery and elimination
Dear Emily
Hope you doing well,
I would like to know the status of your work, if its been resolved then congratulation but if not then according to me you should check the pH at which you are interacting your antibody to gold particle. pH plays a major role for strong conjugation. In most of cases, antibodies binds strongly to 40 nm colloidal gold between pH 7.5 to 8.5 (Based on my experience).
Also avoid the multiple centrifugation steps as you have mentioned in protocol to affect antibody performance and suspend the gold conjugate to similar pH buffer after centrifugation step. Regarding titre, generally the maximum antibody required to saturate the entire surface area of 40 nm colloidal gold of 1.0 ml, approx 3.5 micro gram antibody is required for 1.0 OD gold. (Again, its based on my experiments, publication pending).
You can contact me on [email protected] for any further inquires or looking for custom conjugation services with different size and shape of gold nanoparticles for lateral flow tests.
Good luck for your work.
With best wishes...
hi, i recently came upon this amazingly helpful article about fabrication of AuNP-Antibody nano-conjugates. it specifies the AuNP production method, too. which is very helpful.
hope it comes in handy for you
I have designed using the preincubation format. The sensitivity of detection was much improved, as comparing the common and convention format. The step is easy. Please find the detail from the articles.
Article Preincubation format for a sensitive immunochromatographic a...
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