Preparation and Characterization of Chitosan-Alginate Nanoparticles Conjugated Antibody Payload Anti-EpCAM with BSA and Cell Uptake Test Against Cells T47D
ABSTRACT: Nanoparticle capability in delivering, maintaining the integrity, and conserving the function of protein making it attractive due to the quick emergence of therapeutic protein in the market and also the higher success rate of therapeutic protein compared to small molecule drug. Even so, data shows that the major problem in clinical trial of therapeutic protein includes stability, protein immunogenicity, and side effect elicited by protein-target interaction in the area beyond the desired location. One of the possible way to resolve these problems is by designing a model of nanoparticle-based targeted therapeutic protein delivery. Breast cancer was chosen as the target in the design of targeted therapeutic protein delivery model because of its high prevalence and mortality rate. On the other hand, breast cancer has gene expression characteristic that can be exploited for targeted drug delivery. EpCAM is one of the overly expressed protein in some breast cancer and highly correlated with poor prognosis and higher chance of metastasis. In this study, targeted therapeutic protein delivery model was produced by conjugating anti-EpCAM IgG1 onto chitosan-alginate nanoparticle loaded with BSA as the protein model. Chitosan-alginate BSA nanoparticle was produced by mixing the solution of chitosan, alginate, and BSA with final concentration of 0.33%; 0.133%; and 0.167% respectively. Imaging with TEM shows the resulting nanoparticle having the size varied between 40-200 nm, and shows that the nanoparticle was formed from the aggregation of smaller particle. After conjugated with anti-EpCAM IgG1, nanoparticle size distribution were mostly ranging between 130-245 nm with mean value 184,37 nm, and zeta potential +23,43. Efectivity and selectivity test using T47D breast cancer cell and Vero as control shows the ability of both the unconjugated and conjugated chitosan-alginate nanoparticle in increasing the uptake of BSA-FITC on both cell, but the unconjugated one shows the highest uptake. Conjugation with anti-EpCAM IgG1 AUA1 clone didn�t shows higher selectivity towards T47D. Therefore it is necessary to develop better models using better targeting molecule.
