Project: Portable Aptamer Biosensors for the Detection of Pathogens On-the-Spot (Legionella/Amoeba Complex)

The aim of the project is the development of an on-line miniaturized potentiometric biosensor (a disposable strip) able to simultaneously detect and quantify in an accurate and in an immediate and simple way without the need for trained and skilled personnel the bacterial amount of Legionella pneumophila and of the CO protozoae that can be infected by Legionella pneumophila. Since Legionella pneumophila in certain cases easily can loose its virulence factors or even might die unless it can infect protozoa, detection of amoebae is also of crucial importance in order to prevent legionellosis. Alternatively, the detection of legionella should be directed to virulence linked target structures. Potentiometry was the selected detection method because it is one of the simplest and most cost efficient instrumental techniques for electrochemical sensing. Today, recombinant technologies allow to go beyond the capabilities of animal generated antibodies by introducing molecular designer features into the antibody in the process of its generation. Using in vitro selection and evolution systems and adaption of the Fc moiety, synthetic antibody variants can be generated which are functionally equivalent to IgG molecules but offer a wide range of parameters to be optimised to specific detection problems. The recombinant antibody variant of choice for the proposed projects are Yumabs, a fusion of mouse Fc with scFv-fragments selected from ultra high diversity antibody phage display libraries pioneered by Technische Universität Braunschweig (TUBS). The aim of the project combines the Yumab generation with the biosensoric detection of bound target molecules. _x000D_ Recently, TUBS demonstrated for the first time, that recombinant antibodies could be tailored to the requirements of a particular biosensoric system, instead of other way round. Kinetic binding characteristics of an antibody were tuned to be favourable for a flow through biosensor which finally allowed more than 100 quantitative assays using the same antibody chip. _x000D_ The biosensor for binding of selected pathogen targets is property of and will be developed by another consortium P, Vantix Ltd., UK. (part of the Sensortec Ltd group), a company which has developed an ultra sensitive electrochemical detection technology (patent protected) for the detection of small molecules, proteins, DNA and whole cells. The diagnostic target has been determined by a third P of the consortium, Agrobiogen (ABG), Germany, the CO Participant, which performs a large program directed to diagnosis of animal and human infectious diseases, so far mostly by using PCR, immunolgical and lateral flow approaches for the detection of bacterial and viral diseases. In the past ABG developed a whole clutch of diagnostic applications. One of them was the monitoring of Legionella in different environmental samples. Originally the idea for that was a simple and direct (on-line) or periodic detection. Unfortunately, at that time it was unsuccessfully because of the insufficient sensitivity and robustness of the chosen approach. The only successful approach at that time were PCR based methodologies. Additionally to the detection of Legionella we intend to perform the future assays in parallel to the bacterium also to detect the most common in environmental samples protozoic host genera for Legionella, (Acanthamoeba, Naegleria, and Hartmanella ssp.). The fourth P in the consortium, the University of Veterinary Medicine (VetMedUniVienna), Austria will be responsible for the preparation of all needed kinds of source samples and markers for the generation and selection of Yumabs against Legionella ssp. and their amoebal hosts. ABG will provide all needed PCR assays against the mentioned organisms as Gold standards. Furthermore, ABG will be responsible for sample collection adapters and fittings for the sensor unit. Thus, the consortium consists of four Ps, two SMEs and two universities. Additionally, we invited a minor contractor (LUS) who developed a model of the water unit in the dentist´s chair. This model is a brilliant and outstanding solution for all kind of approvals and validations with the focus to one of the favoured future market nishes for the complex development. The project aims to provide to the industry, governmental institutions and, finally, even interested consumers affordable and easy to use equipment for measuring in different environmental or industrial areas a potential contamination with Legionella and amoeba subspecies. _x000D_ The consortium considers realising the future market access via strategic alliances with large water technology enterprises, leading Test-Service providers and/or In-vitro-Diagnostics producers. The consortium Ps intend to arrange a joint commercialisation strategy. All Ps will benefit from the commercial success of the development depending on their shares in IP, further input in know-how, experience and efforts._x000D_ _x000D_

Acronym APTDETECT (Reference Number: 6994)
Duration 01/08/2012 - 31/07/2016
Project Topic A new approach to detect Legionella and its interrelation with amoebae in water supplies will be developed based on aptamers and screenprinted potentiometric biosensors. The sensor will detect the pathogens on the spot. The core IPs belong to the Ps of the project.
Network Eurostars
Call Eurostars Cut-Off 7

Project partner

Number Name Role Country
5 Technische Universität Braunschweig Partner Germany
5 University of Veterinary Medicine Vienna Partner Austria
5 Agrobiogen GmbH Coordinator Germany
5 VANTIX Ltd. Partner United Kingdom
5 APTARES AG Observer Germany