ERC FUNDED PROJECTS

During the last decade, the principles of biomineralization have increasingly attracted multidisciplinary scientific attention, not only because they touch the interface between the organic/inorganic world but also because they offer fascinating bioinspired solutions to notorious problems in the fields of biotechnology and medicine. However, only one group of animals has the necessary genetic/enzymatic toolkit to control biomineralization: siliceous sponges (Porifera). Based on his pioneering discoveries in poriferan molecular biology and physiological chemistry, the PI has brought biosilicification into the focus of basic and applied research. Through multiple trendsetting approaches the molecular key components for the enzymatic synthesis of polymorphic siliceous skeletal elements in sponges have been elucidated and characterized. Subsequently, they have been employed to synthesize innovative composite materials in vitro. Nonetheless, knowledge of the functional mechanisms involved remains sketchy and harnessing biosilicification, beyond the in vitro synthesis of amorphous nanocomposites, is still impossible. Using a unique blend of cutting-edge techniques in molecular/structural biology, biochemistry, bioengineering, and material sciences, the PI approaches for the first time a comprehensive analysis of natural biomineralization, from gene to biomineral to hierarchically ordered structures of increasing complexity. The groundbreaking discoveries expected will be of extreme importance for understanding poriferan biosilicification. Concurrently, they will contribute to the development of innovative nano-biotechnological and -medical approaches that aim to elicit novel (biogenous) optical waveguide fibers and self-repairing inorganic-organic bone substitution materials.

2 183 600 €

Start date: 2011-06-01, End date: 2017-05-31

Aptamers are single-stranded oligonucleotides which are able to target peptides, proteins, small molecules or live cells by virtue of their well-defined three-dimensional shapes. Aptamers are typically generated by evolution of specific sequences against a given target by in vitro evolution using the process known as SELEX. Progress of this field with respect to drug development has so far been hampered by the relative large size and poor biostability of evolved aptamers composed of unmodified nucleotides, necessitating tedious and extensive post-SELEX truncation and modification approaches. LNA (locked nucleic acid) is a prominent nucleotide modification which is in the process of revolutionizing gene silencing and RNA detection. LNA however has never been included in de novo aptamer evolution. EVOLNA is an ambitious but coherent research program with the objective of transforming the field of aptamer technology. The vision is to enable evolution of aptamers that per se possess most of the desired properties, thereby alleviating the need for extensive post-SELEX procedures. This will be realized by combining the unique properties of LNA with innovative methods for LNA aptamer evolution. LNA aptamer technology is envisioned to enable evolution of aptamers displaying maximum chemical diversity, minimum size and high biostability. The developed strategies will be applicable not only towards evolution of therapeutic aptamers, which will be the main subject of this program, but also towards evolution of aptamers for biosensing, diagnostic and imaging applications. The program is at the very frontier of biotechnology research and spans the areas of chemistry, molecular biology and drug research.

2 497 720 €

Start date: 2011-04-01, End date: 2016-03-31

Biopharmaceutical proteins are typically produced in cultivated mammalian cells, a costly process with limited scalability. Thus products such as monoclonal antibodies are very expensive and often beyond the reach of the world’s poor. The problem is compounded by the fact that important strategies for preventing diseases such as HIV and rabies typically involve large doses of multiple antibodies and other virucidal proteins. Plants have emerged as alternative production platforms for biopharmaceutical proteins because they are less expensive, more scalable and potentially could be transferred to developing countries. Recently, the first products have reached the clinic, but many of them are follow-on products already manufactured in mammalian cells. Here, Prof Julian Ma (St George’s Hospital Medical School, London, UK) and Prof Dr Rainer Fischer (RWTH Aachen University, Germany) aim to develop innovative ways to use plants for the economical, safe and sustainable production of combinations of active pharmaceutical ingredients (APIs) based on recombinant proteins, thereby pushing the boundaries of what can be achieved in plants beyond current capabilities with fermenter-based systems. We will focus on the production of antibodies and lectins against HIV and rabies, with the aim of generating GMP-compliant microbicidal cocktails for evaluation in human trials. Key aspects of the project will include the production of APIs both individually and as combinations in plants, the development of technologies allowing the introduction of transgenes into pre-determined genomic loci, the use of click chemistry to optimize the production and stoichiometry of recombinant protein cocktails, the development of candidate products for both topical and parenteral administration and the development of downstream processing concepts that are transferrable to developing countries, such as minimal processing and processing trains based on pre-assembled disposable modules. We will complete one Phase I clinical trials, each testing a plant-derived product that advances the field in a significant way

3 488 863 €

Start date: 2011-08-01, End date: 2019-01-31

Passive transport through lipid membranes is ubiquitous and fundamental in living systems. The aim of this proposal is to create novel biotechnological tools to study permeation of organic compounds through lipid membranes and protein pores. In particular, I will focus on strategies employed by living organisms to optimize and regulate permeation directly through their membranes. The fundamental principles are probed by creating macroscopic model systems for biological channels and membranes. Simultaneously, new microfluidic tools will allow for a screening of biological relevant organic compounds. Biotechnological experiments investigating permeation of organic molecules into single uni-lamellar vesicles will challenge the dogma of protein controlled membranes transport. Indole, an important signaling molecule for E. coli, is an ideal candidate to demonstrate the feasibility of a novel assay based on a combination of four technologies. Microfluidics provide the controlled environment, holographic optical tweezers confine single vesicles in three dimensions to facilitate ionic current detection and simultaneous auto-fluorescence detection. This unique combination will yield a scalable technology platform to test membrane permeation. However, a deeper understanding of the molecular basis for these passive transport processes is still elusive. Theory predicts that binding potentials for molecules in a protein channel, passive transport can be optimized. Combining microfluidics with holographic optical tweezers provides the optimal means to test this quantitatively. These model experiments will prove that passive transport can be enhanced and optimized by introducing binding sites in protein channels and membranes. Furthermore, the results will guide future design of e.g. antibiotics, DNA vaccines and membrane permeating drugs and fundamentally change our understanding of passive membrane transport.

1 193 759 €

Start date: 2010-12-01, End date: 2015-06-30