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Traditional cancer therapies suffer from toxicity, lack of selectivity, and resistance to drugs. In order to increase the dose reaching the tumor tissue, targeted therapeutic approach is being used. Nanoparticles (NPs) can carry larger amount of drugs, however the amount of these NP systems reaching is less than 10%. Most of these NPs get taken up by cells of the immune system, and segregate in organs of the body such as liver, lungs and spleen. Adding a polymer coating such as Poly ethylene glycol reduces the non-specific uptake. Adding a targeting ligand on the NP surface allows specific interaction with the tumor target and intracellular delivery of the payload. However, the protein coating covers the PEG surface. This study undertakes modification of the antibody of interest (Herceptin) with short PEG moieties. It deals with the characterization of the plain and modified Herceptin and immune response of the mouse macrophage cell line RAW 264.7 to NP-antibody/ NP-modified antibody system. Herceptin and modified Herceptin were characterized using Matrix Assisted Laser Desorption/ Ionization (MALDI) and Mass Spectrometry. Two cytokines, TNF-a and IL-1ß were measured by Enzyme Linked Immuno Sorbent Assay (ELISA). Fluorescent Microscopy study was undertaken. Systematic study of the applicability of the Bradford Assay to the modified antibody was done. Two standard proteins: Bovine Serum Albumin (BSA) and Bovine Gamma Globulin (BGG) were compared with Herceptin response. Modeling of the Herceptin structure was done using the protein sequence available in the Protein databank. |
