"My name is Reka Otvos, I am a PhD student working on the AIMMS PhD project 'Identification of novel bioactive substances on brain receptors'. I am from Hungary, and I obtained my Master's degree in Pharmacy at the University of Debrecen (Hungary). My PhD project is a multidisciplinary project that bridges the Biomolecular Analysis research group and the Department of Molecular and Cellular Neurobiology. Working on a project which bridges two departments is an excellent opportunity to gain experience in different research fields.
My PhD project focuses on the development and application of different hyphenated High Resolution Screening (HRS) methods in order to identify novel bioactive compounds acting on brain receptors. HRS is a technique in which high-performance liquid chromatography (HPLC) separations are coupled to mass spectrometry (MS) detection with in parallel an on-line biochemical assay.
The first part of my project focused on a microfluidic HRS system using nano-LC which allows analysis of biological extracts available only in low amounts, like animal venoms. In the microfluidic HRS system a fluorescence enhancement based bioassay for the Acetylcholine Binding Protein (AChBP) was applied, and used for a snake venom screening campaign. Subsequently, development of a follow-up workflow allowed for MS guided purification and chemical identification of novel bioactives found in the venoms analyzed. Furthermore, the same methodology was applied to the development of a HRS setup for determination of affinity to the 5-Hydroxytryptamine Binding Protein (5HTBP).
To obtain real cellular biological data on bioactive compounds in venoms acting on brain receptors, subsequently an at-line cellular screening methodology was developed with a cell-based functional calcium-flux assay using α7-nicotinic-Acetylcholine receptor over expressing human neuronal cells. This at-line screening system was finally transferred from plate reader format to a real on-line HRS methodology based on continuous cellular infusion with LC eluent followed by an on-line calcium flux assay measured with a flow cytometer. With the HRS screening methods developed new potential lead molecules can be identified from natural complex mixtures for medicines targeting central nervous system (CNS) diseases."