Hey, hi, I am preparing Chitosan-TPP NPs in the ratio 1 (TPP) : 5 (CHI), I am encapsulating DNA inside the NPs and to measure the E.E. I am spinning down the particles and measuring the supernantant (indirect method) unencapsulated DNA, to measure the Encapsulated DNA I need to break the particles without breaking the DNA. I tried using A.A. 2%, didn't work, I tried sonicating with 40 and 60 amplitude, didn't work either. I tried using A.A. 2% + sonicating + 150uL of HCL 2M, this helped to reduce a lot the pellet, but I'm afraid I also destabilized DNA. I need to know a way to break completely the particles without affecting DNA, can anyone happen??? Also, I tried increasing the TPP/CS ratio to 2 : 5 and I have a lot of aggregate, I read I can use tween 80 as a surfactant to avoid aggreation in TPP/CS NPs. Can you tell me if I add tween after doing the particles? With the TPP solution? With the chitosan solution? And in which concentration? Thanks
Also check https://www.frontiersin.org/articles/10.3389/fbioe.2020.00004/full
If you want to break the nanoparticules, you should try with the enzyme chitosanase. And to avoid agregattion it is very important to consider the following factors: what you use to dissolve the complexes. for example dissolved in bidestilated water works better than in PBS, a better size and dispersion. Also take into account the ph of the quitosan and the tpp. What ph do you use your quitosan and TPP?
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