Optimization of PEGylation Conditions for BSA Nanoparticles Using Response Surface Methodology |
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Authors: | Hasan Kouchakzadeh Seyed Abbas Shojaosadati Amir Maghsoudi Ebrahim Vasheghani Farahani |
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Institution: | (1) Biotechnology Group, Chemical Engineering Department, Faculty of Engineering, Tarbiat Modares University, P.O. Box 14115-143, Tehran, Iran; |
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Abstract: | Chemical coupling of polyethylene glycol (PEG) to proteins or particles (PEGylation), prolongs their circulation half-life
by greater than 50-fold, reduces their immunogenicity, and also promotes their accumulation in tumors due to enhanced permeability
and retention effect. Herein, phase separation method was used to prepare bovine serum albumin (BSA) nanoparticles. PEGylation
of BSA nanoparticles was performed by SPA activated mPEG through their free amino groups. Effect of process variables on PEGylation
efficiency of BSA nanoparticles was investigated and optimized through response surface methodology with the amount of free
amino groups as response. Optimum conditions was found to be 32.5 g/l of PEG concentration, PEG-nanoparticle incubation time
of 10 min, incubation temperature of 27°C, and pH of 7 for 5 mg of BSA nanoparticles in 1 mL phosphate buffer. Analysis of
data showed that PEG concentration had the most noticeable effect on the amount of PEGylated amino groups, but pH had the
least. Mean diameter and zeta potential of PEGylated nanoparticles under these conditions were 217 nm and −14 mV, respectively.
In conclusion, PEGylated nanoparticles demonstrated reduction of the negative surface charge compared to the non modified
particles with the zeta potential of −31.7 mV. Drug release from PEGylated nanoparticles was almost slower than non-PEGylated
ones, probably due to existence of a PEG layer around PEGylated particles which makes an extra resistance in opposition to
drug diffusion. |
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