Synthesis of novel phosphorescent oxygen sensing probes and their evaluation by spectroscopy and live cell PLIM microscopy

• Grant number: 19/22578-0
• Support type: Scholarships abroad - Research Internship - Post-doctor
• Effective date (Start): December 16, 2019
• Effective date (End): December 15, 2020
• Field of knowledge: Physical Sciences and Mathematics - Chemistry
• Principal Investigator: Celio Pasquini
• Grantee: Rafael Di Lazaro Gaspar
• Supervisor abroad: Dmitri Papkovsky
• Home Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Local de pesquisa : University College Cork (UCC), Ireland
• Associated to the scholarship: 18/20997-3 - Development of optical oxygen and temperature sensors based on visible and infrared emitting lanthanide ion complexes, BP.PD

Abstract

The research project aims at the development of novel targeted oxygen sensing chemistries and probe structures based on PtPFPP porphyrin scaffold, which provides improved cell penetration and staining and performance in functional live cell analysis, particularly by PLIM microscopy. The new oxygen probes will be synthesized using click modification of PtPFPP to obtain PEGylated derivatives monofunctionalised with NH2 (and possibly COOH) groups, and converting them into reactive N-hydroxysuccinimido and maleimido derivatives which can be used for phosphorescent labelling of protein molecules, such as bovine serum albumin (BSA), transferrin and monoclonal antibodies (e.g. anti-Glut1-mAb) . Structural and photophysical characterization of these intermediates and bioconjugates will be performed, and then they will be tested with cultures of mammalian cells to evaluate their cell staining and oxygen sensing performance. The oxygen images will be obtained on the confocal and wide-field PLIM microscopes installed in the hosting lab. This research will demonstrate the new approach to high-performance oxygen imaging, by combining oxygen probes with improved targeted delivery and selectivity with advanced PLIM based detection of cellular oxygen. It will address the drawbacks of existing oxygen probes and PLIM systems and facilitate their use for live cell and tissue analysis.