ERC FUNDED PROJECTS

Antibody detection assays are used in many fields of biomedicine including the diagnosis of infectious diseases, autoimmune diseases and allergies. Current analytical techniques for antibody detection come with intrinsic limitations such as the requirement for multiple time-consuming incubation steps, multiple reagents, and/or sophisticated equipment. Supported by an ERC consolidator grant we have developed a highly modular sensor concept for antibody-responsive reporter enzymes (AbSens) that addresses many of these challenges. Key advantages include the ability to monitor antibodies directly in solution, easy read-out based on a simple color reaction, adaptability to target any antibody of interest, and high affinity and specificity. We believe that this generic sensor platform could find applications in low-cost point-of-care diagnostics, clinical research, and the development of therapeutic antibodies. The goal of AbSens is to identify those opportunities in the huge market of antibody-based diagnostics where our sensor platform provides unique advantages over existing technologies, both in terms of analytical performance and economics. To enable the next step towards commercialization, the analytical performance of our technology will be compared to current gold standards using relevant clinical samples in collaboration with commercial parties and clinicians. Other commercially important parameters are the long-term stability of the assay components and the development of a yeast-based production system to lower the cost of enzyme production. Based on an in-depth market analysis and the feedback we receive from external stakeholders on the performance of our technology, a realistic strategy will be developed for the further commercialization. In anticipation of exploring the commercialization of our AbSens technology we filed a US provisional patent application in Sept. 2012 on the key underlying technology, which was recently continued via the PCT route.

150 000 €

Start date: 2014-09-01, End date: 2015-08-31

Until now the greatest limitation in the application of bioactive compounds has been the inability of confining them specifically to single cells or subcellular components within the organism. Our recently synthesized photoactive forms of bioactive compounds solve this problem. We have developed effective chemical synthesis methods to attach an azide group to small drug-like molecules, which makes them photoactive. As a result, light irradiation can induce the covalent attachment of these molecules to their target enzymes. By controlling the timing and position of light irradiation it is possible to confine the effect of these molecules in time and space. It is important to emphasize that azidation is the smallest possible modification (adding 3 nitrogen atoms) that makes a compound photoactive and based on our experience it does not alter biological activities of most of the original compounds. Azidated inhibitors give unprecedented freedom to researchers because the covalent compound-target formations allow them to address questions which could not have been addressed before. Three major advantages are obtained by using azidated compounds 1: determination of small molecule interactome becomes highly effective, especially, the weak interactions can be determined, which was not possible before 2: it improves the pharmacodynamic and pharmacokinetic properties of biological compounds as the covalent attachment prolongs their effect. 3: Recently, we showed that photoactivation can be initiated by two-photon excitation, thereby confining the effect to femtoliter volumes and well-controlled spatial locations. This feature provides unprecedented spatial and temporal control in localizing the effect of biological compounds in cellular and subcelluler level in in vivo experiments. By realizing the need for photoactive compounds, the PI has established Drugmotif Ltd., a spin-off company, which provides the customers with special azidated chemicals for high-tech research.

150 000 €

Start date: 2013-12-01, End date: 2014-11-30

Within BMP4EAC we aim to investigate the commercial feasibility of our newly discovered and highly specific antibodies against BMP4 and one of its receptors, BMPR1a, for imaging applications in oncology. BMP4 and BMPR1a are highly expressed in esophageal adenocarcinoma (EAC) and other tumors as well as their metastases. The specificity, strong binding capacity, rapid clearance, high tissue penetration level, and flexibility of our antibodies is unprecedented and makes them highly suitable for in vivo imaging applications. The opportunity: The current methods for evaluation of disease stage consist of diverse modalities, including, CT and PET-CT scans, and ultrasonography. These techniques have major limitations to accurately detect metastasis and are inadequate for monitoring disease response. In the clinical setting we foresee applications of our proprietary technology in the non-invasive diagnosis of tumors and metastases with high expression of BMP4 and/or BMPR1a (e.g. EAC), identification of patients with high chance to respond to BMP4 inhibitors, follow tumor progression during treatment, and facilitated localization of small metastases during surgical treatment. Furthermore, the labeled antibodies can be used to investigate the efficacy of novel therapeutic agents by following tumor progression in animal models in a research setting. The project and expected outcomes: Within the ERC PoC we will explore the commercial feasibility by in vivo validation experiments as well as by performing essential research for the formulation of a business proposition, strengthening our IP position, and developing a sound business plan. These activities will result in a proposition package that will be used to present the commercial potential to investors and other strategic partners to attract funding after completion of the ERC PoC and potentially even initiate licensing and partnership deals.

149 840 €

Start date: 2014-09-01, End date: 2016-02-29

Assembly of combinatorial genetic libraries for identification of biomolecules with novel or improved properties requires high fidelity and efficiency to produce the greatest spectrum of genetically diverse clones. We have been developing homologous recombination (HR) as a platform for the production of combinatorial genetic libraries for affinity maturation and diversification of human therapeutic antibodies. Therapeutic antibodies have a great clinical potential in various therapeutic settings including the treatment of a number of oncology, autoimmune and infectious diseases, organ transplantation, and others. In brief, we describe a method, where CDR-encoding DNA oligos and a gapped vector containing the heavy and light chain genes are cotransformed into budding yeast Saccharomyces cerevisiae for in vivo assembly by HR. Importantly, the affinity of resulting antibody clones in the generated library can be directly assayed by yeast surface display without subcloning and retransformation. Furthermore, mating two haploid yeast strain libraries each encoding a variation of heavy chain or light chain genes enables fast screening of the heavy/light chain combinations displayed by the resulting diploids. Finally, this method can be generalized to generate combinatorial genetic libraries for other applications.

150 000 €

Start date: 2014-05-01, End date: 2015-04-30