I'm trying to attach an antibody with this method but I see that after of some minutes the nanoparticles change the color from the original color to "clear" color and in the UV/vis the plasmon resonance is not shown. My nanoparticles are stabilized with alginate.
Could you please tell me where does the color come from? Is it from the drug, if yes then what kind of drug is it (according to chemical structure classification)?
From what I know, EDC only enhance the formation of peptide bond. So most likely the loss of color would be due to the break of the chromophore because of the reaction with NHS. But there are many possibility that could happen.
Radif
Thanks.
The color comes from the nanoparticles, when I add EDC-NHS the color disappear.
You said that it is stabilized with alginate, could you explain the whole component of your nanoparticle? Because as far as I know alginate is colorless when dissolved and cloudy (white) when became nanoparticle.
I'm working with nanoparticles for enzymes immoblilization - EDC-NHS usually and nothing wrong was happened. What type of nanoparticles were used in Your research? I think it is important becauseNHS and EDC are highly reactive with functional groups etc.
It will depend on the linker species you have attached to the nanoparticle surface and the concentration of EDC.HCl and sNHS which you are using in order to activate the moiety for peptide bond formation. You may need to further stabilise your nanoparticles with a polymer such as PEG to prevent aggregation of the particles.
Thank you all!!... I use the alginate as stabilizing agent, I saw that this polymer is used for having a coupling using EDC/NHS so I thought that this polymer could be a good chemical, for having the coupling with the antibody and also the stabilization of nanoparticle.
Dear Valdez.. would let me know which polymer you are using .. u had stabilized with alginate.. are u using polymer nano particle?? and what pH u are using for conjugation.. usually EDC NHS chemistry will be active only when pH below 7.4..
I agree with all above folks, specially, Chandan. I would suggest you to check pH of alginate particle solution before and after addition of EDC/NHS, to make sure the coupling pH (6~7) to be adjusted.
I would suggest you that, there is no direct impact of colour change but when you conjugate a molecule by EDC/NHS method generally particle size increase and and there PH plays a major role in colour change.
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