MINIATURIZATION OF MICROARRAY BASED SCREENING OF HSP INHIBITORS USING NANOPARTICLES AND SYNTHESISING NANOCARRIER FOR INHIBITORS

Abstract

Folding of protein is carried out by molecular chaperones like Heat Shock Proteins (HSP), out of which HSP70 and 90 are most vital members. Expression of HSP70 and 90 surges by a variety of cellular stresses. Human cancer cells display comparable stress symptoms. On a molecular level, this leads to a surplus of HSP in cancer cells, especially HSP90. HSP has ATP dependent protein folding action. Small molecules can interact with HSP by binding to nucleotide binding site, thus hampering folding of proteins. This leads to the breakdown of many activities in oncogenic cells. Few molecules have been recognized, that attaches to the ATP binding site. Microarray systems can be used to test in-house library of new inhibitors of HSP. Q-Dots have interesting electrical and optical properties. Exploitation of its surface plasma resonance effect for optical sensing plays a momentous role in the biomolecular diagnostic assay. Cancer nanotherapeutics are rapidly developing and being applied to solve several restrictions of traditional drug delivery systems such as nonspecific binding, low bioavailability, reduced solubility in aqueous media, truncated therapeutic indices and generalized biodistribution. The nanoparticle is in optimum size range and has unique surface characteristics which lead to prolonged circulation time in blood. They serve as carriers to drug molecules with high specificity for tumours, because of their enhanced permeability, amplified retention effect, and microenvironment of tumour cells. This thesis work aims at miniaturizing the microarray assay using Streptavidin conjugated Quantum-Dots. Q-Dots are spotted on Nitrocellulose surface and quenching of their signal by HSP attached to biotinylated-ATP so that it can be further used for screening inhibitors that bind to nucleotide binding site of HSP. It also aims at synthesizing HSP inhibitor loaded niosomes that can be used for targeted delivery to cancer cells