Automation for Life Sciences - Institute for Automation, University Rostock

Abstract

Rostock, with its nearly 200.000 inhabitants, is the biggest city in Mecklenburg-Vorpommern. It is easy to reach over the highway in about two hours north from Berlin or east of Hamburg. Since 2002 the direct connection to Munich from the airport of Rostock makes it possible to reach Rostock from every point in the world. The town is directly situated at the Baltic Sea and thus the main tourist center in North Germany. Before the political changes in East Germany Rostock was well-known for it’s shipbuilding industry and had a great port with millions of tons stock turn every year. Today most of the heavy industry is gone and Rostock has developed itself to a progressive city blossoming with high-tech industry. The strong connection with the University of Rostock (founded in 1419) is the basis for the developments in high-tech. Different technology centers have been developing over the last years, one of these is the technology park in Warnemuende. The Institute of Automation (IAT) is situated in the technology park Warnemuende. It started in 1994 with 13 people in research and education and has now 52 people working in different fields of measuring techniques, control and automation. The institute is working successfully in different projects in strong connection to the life science industry. Different companies and non-profit organizations have been founded out of the institute such as the Institute for Measuring and Sensor Systems (non profit organization working in the field of applied research), the Analytical Instrument GmbH (customer specific solutions in system integration and system development) and the Telematika GmbH (IT solutions). The project and industry money for 2001 was more than 7 million DM. About three years ago the professorship for laboratory automation (the first in the world) at the institute for automation was offered to Dr. Kerstin Thurow. With her 29 years she was the youngest university professor in Germany and one of the very few woman professor in engineering sciences. With her background in chemistry (PhD 1995) and engineering sciences (habilitation 1999) she had already been working for many years at the border between chemistry and engineering. Since 2000 she is also the managing director of the institute. The institute of Automation is developing customized solutions in the field of life science automation. Life sciences is for us not only drug screening and high throughput screening but also means more than the handling of microtiterplates. Within the last years we saw a dramatic increase of automation and HTS needs in different fields of chemical and pharmaceutical industry such as catalysis research, synthesis, synthesis optimization, product control, screening of microorganisms, etc. The main goal is not always the handling of as much samples as possible. The great challenge in our work is the fact that we have customers from different companies with very specific problems. Every solution is customer specific with a high amount of engineering work. Most of our customers come to us after they failed in finding commercial companies on the market who are able to solve their problems. “Our big advantage is the interdisciplinary team. In the team electrical engineers, IT-people, mechanical engineers, chemist and biologists are working together. This gives us the chance to understand the complex needs of our customers and to offer complete solutions from the planning over the development to the application of the systems” says Kerstin Thurow. The key competences of the institute include:•Instrument design / Reaction technology•System development•Robotics•Automated Analyses including preanalytical sample treatment•Remote Control•HTS•Process Control Systems•Internet LIMS The main focus of our work is the development of complex systems for different applications in life sciences. Companies like Beckman, Zymark, CRS or Tecan provide complex solutions using robotic systems. These solutions often do not meet all requirements of customers. Customer wish to have specific devices integrated in their system or have special automation needs which can not be solved with commercially available systems. Thus the system integration is one of the rapidly developing fields in laboratory automation. In Germany the IAT is working as a system integrator for ORCA robotic systems (Beckman Coulter). Customer specific integrations can be done with any device for sample preparation or analysis. Devices which have been recently integrated include the Rotanta centrifuge (Hettich), the NovoStar (BMG) as an alternative for fluorescence measurements, microtiterplate shakers (H+P), heated MTP shakers (Eppendorf) or Ultravap (Zinser) as evaporator. The integration of the systems requires hardware as well as software developments. Beside the hardware interfaces often additional mechanical reconfiguration of the system is necessary to ensure the direct access of the robot to all samples and stations. The robot control software SAMI NT is used as a graphic object oriented interface to enable the control and scheduling of the additional instruments via the robot control software. Currently the integration of new centrifuges as well as the integration of specialized shakers with incubation or evaporators are under development. Beside the integration of single devices we also build complex robotic systems. Usually ORCA (Beckman-Coulter) or Zymate (Zymark) are used as robots, but for specific customer needs the integration of other robots is possible too. Typical applications in the biological section include enzyme inhibition tests for biological activity testing or fluorescence detection with cell cultures (e.g., determination of Ca2+ changes in dependence on drugs). Whereas there is still a big orientation of companies like Beckman, Zymark, Tecan or Cybio to the automation with microtiter plate formats we see an increasing interest in additional techniques. Chemical processes often do not require an ultrahigh throughput, normally they use more material in order to ensure the upscaling of the tested processes and typically work in the single flask format. Complex robotic systems are built at the IAT for applications in product control in chemical production, synthesis optimization or metabolite screening for drug testing. These systems include a single sample handling with variable volumes from 1 ml to 250 ml. Specialized devices such as balance, screwer, dilutors or solid dispenser have to be integrated for these systems. The degree of automation depends on the specific applications. Not always complete solutions with complex robots are necessary and economic. For sample preparation procedures (liquid handling) and direct connection to analytical devices xyz-robots are used at the IAT. These robots are based on a CTC CombiPAL system and have been combined with various special devices such as heaters, stirrers, filtration or SPME units, dilutors and injection ports. The sample preparation task allows to apply user specific methods in sample preparation. Sampler and chromatographic systems communicate for transmitting remote START/STOP- information via hardware handshake lines. Because of the flexible system strategy, the system can be set up easily for different applications. The developed systems can be sold to industry partners. On the other hand small biotech and life science companies can rent the systems for a specific time to do their own screenings. Thus a team of chemists and biologists is using the systems for application development in different fields. The third aspect of using complex robotic systems is user training. Since working as system integrators we prefer to have commercially available systems which can be integrated into complex systems. Sometimes the requirements of the customers do not match to the possibilities of available devices; in this cases we are trying to develop appropriate systems. Typical developments are in the field of reaction systems. Reaction systems in catalysis research or drug refinement require often heavy reaction conditions such as high temperatures up to 200°C and high pressure up to 200 bar. Thus the development of a suitable system was done at the IAT1. The multi parallel reactor system can handle a flexible number of up to eight independent single heated reactors (2 ml volume) which can be used at different pressures up to 200 bar. Up to three reactor systems can be coupled together. A complex reactor system for volumes up to 500 μl which can be integrated into robotic systems is currently under development. Other developments were done for automatic refrigerators (down to –50°C) which can be operated by an ORCA robot. This development was necessary due to the requirements of storage of microtiter plates at low temperatures to ensure the breakdown of the cell material. The use of combinatorial methods in chemistry and life science has been developing rapidly within the last years. One of the “bottlenecks” in synthesis control in pharmacy and life sciences is still the characterization and speciation of biotechnological reaction products. Often, prior to analysis, a preanalytical treatment of the samples is necessary, especially for complex matrices like synthesis solutions which may contain solids or byproducts. Classical analytical systems for HTS applications include mostly spectroscopic methods such as fluorescence or absorbance reader. These systems are highly parallel and can measure numerous sample within a short time. The disadvantage of the existing systems is the applicability to a limited number of application (such as luminescence-based ELISA, Immunoassays or enzyme kinetics) and their disability for structure elucidation. Classical analytical methods such as GC, LC or CE in combination with specific detection methods (e.g., mass spectrometry) can solve these problems. Due to the often long analyses times the throughput of these methods is very low and does not meet the requirements of automated robotic systems. The IAT is thus developing new methods for rapid analysis of complex mixtures. Shorter analyses times can be realized with fast chromatographic methods. The use of fast GC in combination with rapid screening mass spectrometers (e.g., TOF) can reduce analysis times to less than two minutes. On the other hand the institute is developing methods for complex solutions without chromatographic separations. The selectivity is then included in the used analytical method. For the determination of enantiomeric excess we use specific derivatization procedures with mass tagged chirale agents. As a result of the kinetic racemate separation we are able to determine the ee of synthesis solutions within two minutes maximum. The other possibility is the use of high resolution mass spectrometry for the exact mass detection of different compounds in complex reaction mixtures. The coupling of FTICR/MS with an CTC robot for sample preparation and introduction ensures short analysis times and thus the required throughput for the majority of chemical and catalysis applications. Additionally software solutions for automated data analyses (e.g., calculation of conversion, recovery, ee from chromatographic data or calculation of element composition from exact masses from FTICR/MS) are developed at the IAT. To ensure fully automated systems with 24/7 availability possibility all analytical devices are directly integrated into robotic systems. The Institute for Automation is working already for many years as an application and demonstration laboratory for Agilent analyses devices. Agilent systems are usually used for method development and integration into complex systems, but other systems can be integrated, too (e.g., HPLC - Merck-Hitachi; Quattro II - Micromass or Apex III - Bruker). Another important field of work is the development of Laboratory Information Management Systems (LIMS) which really matches the needs of the customers. Many companies offer customers specific solutions including experiment planning and raw data archives. The main research focus of the IAT is the connection between LIM systems and process periphery. This direct coupling of management systems to the used devices (reactors, robots, analysis devices) is still the missing link. Technically a uniform interface can be used to integrate any system. All available systems are coupled to the LIMS as a “Virtual Lab”. Password protected access allows the user remote access to the machines and the data. All robotic systems are equipped with cameras for direct control of the system operation. In the case of errors, messages to cell phones or emails are generated to inform the operators. With the integration of the process level a complete automated system is available which includes experiment planning, operation of all devices and systems, data storage and automated generation of secondary data, which can be stored in data bases. The strict use of internet technology allows an easy service and extension for the system as well as low costs for different clients. Every system is set up customer specific but due to its “open” structure it can be easily reconfigured and extended by the user. The Institute for Automation is not a typical institute at a German university. It represents a modern innovative new way of successful cooperation between industry and universities. The industry has a great demand in developments of automated solutions in different fields of life science applications. The universities have the requirement to find new interesting research topics to ensure a state of the art education of the students. Thus a perfect combination of interests from both sides is possible. The director of the IAT runs the institute like a company: developments should lead to a product and should not be more expensive than the money we can get for them on the market. We feel responsible for our scientists and technicians and would like to offer them future work and want to save and expand working places in the region. Different big projects connected to life science engineering and automation are currently established at the IAT. This include:•Combinatorial methods for the efficient development of catalysts (BMBF / industry)•Life Science Catalysis and Engineering (BMBF / industry)•New drugs and materials - screening procedures and product development (EU / industry)•High performance Catalysis and Analysis (BMBF)•Competence Center for Life Science Automation (BMBF / industry)•Numerous bilateral projects with partners from chemical and pharmaceutical companies The big advantage for the future is being in a niche market which can be defined as complete customer specific automation. Their are still a high number of unsolved problems for different fields of application. New developments include reaction systems for catalysis research, sampling from reactors, systems for high throughput applications in synthesis, the handling of microorganisms or cell culturing procedures and other more. A new building with even more high specialized laboratory space (especially chemical and biological labs) will be available in 2004. Next year the IAT plans the foundation of a new company for high throughput analysis. And this will probably not be the end of future developments.