computer controlled automated assay for comprehensive studies of enzyme kinetic parameters计算机控制自动分析综合酶动力学参数的研究.pdfVIP

Computer Controlled Automated Assay for Comprehensive Studies of Enzyme Kinetic Parameters 1. 1. 2 1 1 1 Felix Bonowski , Ana Kitanovic , Peter Ruoff , Jinda Holzwarth , Igor Kitanovic , Van Ngoc Bui , Elke 1 ¨ 1 Lederer , Stefan Wolfl * 1 Department of Biology, Institute for Pharmacy and Molecular Biotechnology, Ruperto-Carola University of Heidelberg, Heidelberg, Germany, 2 Centre for Organelle Research, Faculty of Science and Technology, University of Stavanger, Stavanger, Norway Abstract Stability and biological activity of proteins is highly dependent on their physicochemical environment. The development of realistic models of biological systems necessitates quantitative information on the response to changes of external conditions like pH, salinity and concentrations of substrates and allosteric modulators. Changes in just a few variable parameters rapidly lead to large numbers of experimental conditions, which go beyond the experimental capacity of most research groups. We implemented a computer-aided experimenting framework (‘‘robot lab assistant’’) that allows us to parameterize abstract, human-readable descriptions of micro-plate based experiments with variable parameters and execute them on a conventional 8 channel liquid handling robot fitted with a sensitive plate reader. A set of newly developed R-packages translates the instructions into machine commands, executes them, collects the data and processes it without user-interaction. By combining script-driven experimental planning, execution and data-analysis, our system can react to experimental