full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
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Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (2) 1084-1097 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1090found released at pH 5.6, whereas 63.5% <strong>and</strong>96% <strong>of</strong> insulin was released in pH 7.4 <strong>and</strong> 1.2media, respectively.Effect <strong>of</strong> Method <strong>of</strong> preparation on InsulinIntegrity <strong>and</strong> Bioactivity : The integrity <strong>of</strong> insulinduring the preparation <strong>of</strong> nanoparticles (organicsolvent, homogenization, surfactants, freeze dryingsteps), was evaluated by extracting insulin fromnanoparticles <strong>and</strong> analyzing the molecular weightby MALDI-TOF Mass Spectrometry. The massspectra showed only a molecular peak at m/z2866.4 (doubly charged, molecular weight <strong>of</strong>bovine insulin is 5733.5 Da) for samples preparedby both methods as well as the insulin solution(Fig. 4). This indicated that insulin retained itsintegrity <strong>and</strong> insulin was not degraded in thepreparation process in all the methods evaluated.The immunogenicity <strong>of</strong> insulin afterencapsulation into nanoparticles by differentmethods was evaluated using ELISA test. Thedata showed that the antigen activity <strong>of</strong> insulinremained particularly unaffected by DCMTable 2: Effect <strong>of</strong> tween 20 concentration onimmunogenicty <strong>of</strong> insulin. Data represented as themean+SD. (n=3).Tween 20Concentration (%) 0 1 2 4Antigenicity (%) 23.11+0.02 29.51+0.72 27.61+0.16 28.18+13.38(102.46% + 19% ), or ET-AC (97.63%+ 18.18%)in ESE method. However, only 28.18+13.38%insulin antigenicity was retained fromnanoparticles prepared by MM method (Fig. 5).Tween-20 <strong>and</strong> MPEG-350 were used inthe preparation <strong>of</strong> insulin nanoparticles using MMmethod. To differentiate which <strong>of</strong> these twosurfactants reduced antigenicity <strong>of</strong> insulin, ELISAassays were conducted using nanoparticlesprepared at different Tween-20 concentrations.The antigenicity <strong>of</strong> insulin remained constant inthe Tween-20 range <strong>of</strong> 1%-4% while it isFig. 6. Cell Viability (%) in the presence <strong>of</strong> insulin solution<strong>and</strong> insulin nanoparticles. Data presented as the mean+SD.(n=3).significantly reduced (P