Micro-spectroscopic studies on Mycobacteria & Cancer cells

Abstract

Evasion of the immune system is a key hallmark of cancer progression. Developing strategies that diagnose early the immune-cancer synapse may lead to long term cures from these aggressive cancers. We aimed to develop optical endoscopic Raman imaging methodology to identify specific regions in the cancer-immune synapse currently beyond detection in vivo and improve survival of cancer patients. When these methodologies are made simpler and direct, success of implementation increases. This early stage detection reduces burden on both patients and physicians for better treatment management. Chapter 6 aimed in designing antibodies for cancer detection which can be applied for Raman Imaging. Instead of designing complete antibodies we aimed in designing recombinant antibodies namely single chain variable Fragment (scFv) to target Agr2/3 on cancer cells. ScFv hold many advantages than the complete antibodies. The problem was addressed in two different ways. (i) Imaging of Agr2/3 using Recombinant Antibodies: GFP labeled recombinant antibodies (scFv4) against Ag2/3 were designed and expressed in breast cancer cell line. Fluorescence live cell imaging was performed. After expression it was blotted against CD20, GFP and Agr2 to access their affinity towards the target. (ii) Purification of scFv7 for Agr2 identification-Towards Dipstick technology: scFv7 was expressed in bacteria and purified from them to make it identify from body fluids. scFv7 binding with Agr2 was analyzed using Elisa. Chapter 7 summarizes the thesis and brings out the key concepts by which the current study can be extended to diverse applications. Identification of bacterial infection and cancer are few of the challenges the posted by health care sectors. We hope in the near future that the vibrational microspectroscopic techniques like IR and Raman may transcend the current diagnostic methods and emerge as a rapid, sensitive and specific tool. Moreover, with the advent of many machine learning methods these techniques can be close to complete automation in diagnosis and screening programs which reduces the burden on technicians and physicians who have to handle large amount of samples in countries like India