Highest productivity in drug discovery experiments

Dr. Bernhard Ronacher

“Working with traditional array-based systems was quite frustrating. Complex microarray handling with various manual processes, lack of robustness, and no possibility to run kinetics did not lead to satisfactory results. Therefore I developed the hybcell technology with the objective to eliminate these limitations. I wanted to provide a highly productive system which generates superior quality results, and is easy to use”
-Dr. Bernhard Ronacher, inventor of the hybcell and co-founder of Anagnostics Bioanalysis

Multiplexed technologies increase productivity, if two conditions are met: there is no room for tedious manual labor and derived data offers additional content. Anagnostics' technology, based on a cylindrical microarray, processes up to 96 assays fully automated in a single device. The system's unique capability of kinetic scans is applied to on/off rate measurements or activity determinations of enzymes.

Anagnostics Bioanalysis is a Hightech-Startup, based in Austria, which specializes in developing microarray based systems and assays for research and routine diagnostics. Founded in 2005, Anagnostics has developed the unique cylindrical microarray, the patented hybcell, and built a completely automated system, the hyborg.

Easy handling and automated workflow
The design of the hybcell allows for great versatility, if required. However, once an assay is established, the whole process is standardized and easily performed in any laboratory.

The following work flow of a typical assay demonstrates the ease of use:

Step 1: Select and apply detector molecules onto the hybcell surface:
Detector molecules (DNA or proteins) are immobilized (printed) on the pre-treated surface of the inner cylinder (black). This inner cylinder is placed into a cylindrical container (transparent). Together these parts form the hybcell. Printing is done by Anagnostics as a service.

Step 2: Add samples:
Liquid sample is filled into the hybcell via the axial central channel with help of a pipette or robot. The hybcell is closed with a rubber lid by the user.

Step 3: Link sample and hybcell ID and transfer hybcell to hyborg:
Each sample is linked to a uniquely barcoded hybcell. After transferring the hybcell to the hyborg no more manual labor is necessary – hyborg operates fully automatic.

Step 4: Process samples:
Within the hyborg one hybcell after the other is transferred into the hybridization and analysis chamber. By lowering the inner cylinder sample liquid is rising and the annular ring of sample liquid is forming over the printed detector molecules. At this point in time interaction of sample and detector molecules starts.

One step after the other of the pre-defined process queue is executed by the hyborg. Possibilities range from temperature profiles, exchange of liquids and multi-color fluorescence scans or simple programming of rotation speed of the inner cylinder.

Step 5: View and confirm results:
Results are provided both in tabular and graphic format. Each result is stored within a database and can be exported or printed.

Unique cylindrical microarray, the patented hybcell.

Productivity through automation: hyborg system
Anagnostics' hyborg is a fully automated and integrated device for processing and analyzing cylindrical DNA and protein microarrays (hybcells). hyborg integrates thermocycler, hybridization station and scanner in a single device. hyborg's versatility makes no difference between DNA or protein microarray applications. All processing steps – from temperature profiles or exchange of liquids to multi-color scanning – are freely configurable by the scientific user and controlled and documented by a PC. Please refer to image 1 in the image gallery at the bottom.

Hyborg's concept of totally integrated automation guarantees maximum reproducibility of assays. Autonomous processing of up to 96 hybcells reduces hands-on-time to a minimum and provides maximum ease of use. Seamless electronic documentation of operation and states fulfills highest quality assurance requirements. In short, hyborg offers:
− Derive (binding) kinetics by analyzing consecutive scans
− Computer-controlled temperatures, exchange of buffers, scans
− Pre-assigned and thus reproducible starting temperature
− Robust hybridization due to rotational sample agitation
− Highly reproducible timing
− Fully automated handling of up to 96 samples

Intuitive software tools
Efficient operation of any lab equipment is key in today's SW intensive lab routine. Ronacher explains:
'While we wanted to maintain the utmost flexibility of the system we did not want to burden lab technicians with complex software tools. As a result we developed two software interfaces, one for the assay developers, hybwiz, and one for system operators, hybcon. That way the scientist or operator can focus on the task at hand. Both solutions do integrate well, of course. As a general assay development tool, hybwiz can be downloaded free of charge from the Anagnostics website.' Please refer to images 2 and 3 in the image gallery at the bottom.

Application focus: assays for drug discovery
Due to the technology's flexibility the application areas are virtually unlimited. However, in the research application space Anagnostics' focus lies on kinetic protein applications, such as multiplexed measurement of on/off rates in protein-antibody binding experiments (e.g. in biomarker studies) or kinetic measurement of protease activity. The use of laser fluorescence detection provides highest detection selectivity. Please refer to images 4 and 5 in the image gallery at the bottom.

DNA applications focus on determination of kinetics like melting curves and on-spot reactions (primer extension,...). A 'PCR-like-protocol' is in preparation, aiming to combine PCR and microarray hybridization into a single test. This protocol will dramatically reduce complexity of many DNA microarray applications. A number of technical notes can be accessed on Anagnostics' website. Please refer to image 6 in the image gallery at the bottom.

Outlook
Having designed hybcell/hyborg for routine drug discovery applications Anagnostics is also developing assays for clinical use.

Contact details:
Anagnostics Bioanalysis GmbH
A-4020 Linz/Austria
T: +43 732 9015 6080
E: info@anagnostics.com
www.anagnostics.com