Multiplexed assays enter drug discovery labs

By Bernhard Ronacher, Anagnostics Bioanalysis GmbH

How multiplexed technologies can increase productivity if they produce reliable, consistent data and eliminate tedious and error-prone manual work in the laboratory.

Anagnostics Bioanalysis GmbH is a high tech startup based in St Valentin, Austria, which specialises in developing microarray based systems and protein and DNA assays for research and routine diagnostics. Founded in 2005, Anagnostics has developed a unique cylindrical microarray, the patented hybcell, and built a completely automated system around it, the hyborg.

Dr. Bernhard Ronacher, inventor of the hybcell and co-founder of Anagnostics Bioanalysis, points out: "Working with traditional array-based systems was quite frustrating. Complex microarray handling with various manual processes, lack of robustness and the lack of kinetics measurements did not lead to productive work and satisfactory results. Therefore Anagnostics developed the hybcell technology with the objective to eliminate these limitations. We wanted to provide a highly productive system which generates superior quality results, and is easy to use."

Easy handling and automated work flow

The design of the hybcell allows for great versatility, if required. The specially developed  surface of the cylindrical microarray allows binding of DNA as well as proteins, which is a significant difference to most other multiplexed or microarray systems on the market. However, once an assay is established, the whole process is standardised and easily performed in any laboratory by standard technical personnel.   

Anagnostics' hyborg system is a fully automated and integrated device for processing and analysing the cylindrical hybcells. The hyborg integrates sample loading module, microfluidics module, thermocycler, hybridisation station and fluorescence scanner in a single device trough automation. All processing steps - from temperature profiles or exchange of liquids to multi-colour scanning - are freely configurable by the scientific user and controlled and documented by the hybwiz and hybcon software platforms.

Intuitive software tools

Efficient operation of any lab equipment is key in today's data intensive intensive lab routine. Anagnostics maintained utmost flexibility of the system but did not want to burden lab technicians with complex software tools. As a result, two software interfaces were developed, 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 seamlessly, of course.

DNA and protein 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, high throughput selection of antibodies based on affinity, antibody QC) or kinetic measurement of protease activities.

The use of fluorescence detection with specific fluorophores provides highest selectivity  at high sensitivity. Even high throughput experiments can be conducted completely automated through the combination of the 4.000- spot hybcell surface and the 96 sample capacity, a completely unique capability.

DNA applications focus on the determination of DNA melting curves and on-spot reactions (primer extension, compact sequencing, nano-preparative DNA target enrichment). A 'PCR-like-protocol' has been developed, aiming to combine a two step PCR and microarray hybridisation process into a single assay, but in a highly multiplexed manner. This protocol dramatically reduces the complexity and required time of many DNA microarray applications.

The time required for one complete multiplexed assay ranges from 10 to about 90 minutes.

A number of technical notes describing the developed assays and applications can be accessed on Anagnostics' website.

Outlook

Anagnostics developers have successfully worked with a number of partners both in academic institutions as well as in pharmaceutical drug discovery laboratories to prove the hybcell technology's capabilities and the performance of the hyborg system. This new and revolutionary hyborg system for multiplexed DNA and protein assays is now commercially available.

Dr Bernhard Ronacher, a molecular biologist, geneticist and biochemist, developed the hybcell microarray technology. He headed the molecular diagnostics laboratory of Lambda GmbH, where he established the DNA chip and array technology and in 2001 the first Austrian biochip, ParoCheck, was filed for patent. At the university, he uncovered  the course of infection by Rhino viruses, responsible for the common cold. His work was fundamental in the development of anti-viral agents and was rewarded by the European Hoechst Foundation.