I made HSA nanoparticles by desolvation method. I took 180mg HSA and dissovled it in 4mL of milliQ water (45mg/ml) and then in the protien solution added 2.5mL of ethanol dropwise, then these nanoparticles were stablized by adding EDC (5mg in 0.5ml of milliQ). The solution was left for 3hrs for nanoparticles synthesis under constant stirring at 1250. The nanoparticles size was checked by zeta sizer and the size found to be 64nm. Then i tried to settle down the nanoparticle at 13000rpm for 10 min but I'm not getting pellet. Further I also tried to increase the cetrifugation speed and time still i did not get the pellet. Any suggetion about this query would be appreciated.
Hi Anuradha Mishra
you can also use the temperature here. try 4° 13000 rpm for 30 to 35 minutes. keeping low temperature during centrifugation can help in getting the precipitate.
With such a small hydrodynamic diameter (less than 100 nm) you may need more centrifugal force to sediment the nanoparticles. When you say 13000 rpm, you should convert your value to "g", because, depending on the centrifuge, size of rotor, type of rotor (vasculating or fixed) the conversion between rpm and g is different. I would try to centrifuge up to about 20000xg and maybe add NaCl (approx 10 mM if it does not influence the formulation) to improve sedimentation.
If you can't reach that centrifugal force with your equipment, you can try to purify by ultrafiltration, centrifuging with filters of about 3KDa pore diameter.
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