A multi-Fc-species system for recombinant antibody production

<p>Abstract</p> <p>Background</p> <p>Genomic, transcriptomic and proteomic projects often suffer from a lack of functional validation creating a strong demand for specific and versatile antibodies. Antibody phage display represents an attractive approach to select rapidly <it>in vitro </it>the equivalent of monoclonal antibodies, like single chain Fv antibodies, in an inexpensive and animal free way. However, so far, recombinant antibodies have not managed to impose themselves as efficient alternatives to natural antibodies.</p> <p>Results</p> <p>We developed a series of vectors that allow one to easily fuse single chain Fv antibodies to Fc domains of immunoglobulins, improving their sensitivity and facilitating their use. This series enables the fusion of single chain Fv antibodies with human, mouse or rabbit Fc so that a given antibody is no longer restricted to a particular species. This opens up unlimited multiplexing possibilities and gives additional value to recombinant antibodies. We also show that this multi-Fc species production system can be applied to natural monoclonal antibodies cloned as single chain Fv antibodies and we converted the widely used 9E10 mouse anti-Myc-tag antibody into a human and a rabbit antibody.</p> <p>Conclusion</p> <p>Altogether, this new expression system, that brings constant quality, sensitivity and unique versatility, will be important to broaden the use of recombinant and natural monoclonal antibodies both for laboratory and diagnosis use.</p

Tissue-specific targeting of DNA nanodevices in a multicellular living organism

Nucleic acid nanodevices present great potential as agents for logic-based therapeutic intervention as well as in basic biology. Often, however, the disease targets that need corrective action are localized in specific organs and thus realizing the full potential of DNA nanodevices also requires ways to target them to specific cell-types in vivo. Here we show that by exploiting either endogenous or synthetic receptor-ligand interactions and by leveraging the biological barriers presented by the organism, we can target extraneously introduced DNA nanodevices to specific cell types in C. elegans, with sub-cellular precision. The amenability of DNA nanostructures to tissue-specific targeting in vivo significantly expands their utility in biomedical applications and discovery biology

Specific GFP-binding artificial proteins ( Rep): a new tool for in vitro to live cell applications

International audienceA family of artificial proteins, named αRep, based on a natural family of helical repeat was previously designed. αRep members are efficiently expressed, folded and extremely stable proteins. A large αRep library was constructed creating proteins with a randomized interaction surface. In the present study, we show that the αRep library is an efficient source of tailor-made specific proteins with direct applications in biochemistry and cell biology. From this library, we selected by phage display αRep binders with nanomolar dissociation constants against the GFP. The structures of two independent αRep binders in complex with the GFP target were solved by X-ray crystallography revealing two totally different binding modes. The affinity of the selected αReps for GFP proved sufficient for practically useful applications such as pull-down experiments. αReps are disulfide free proteins and are efficiently and functionally expressed in eukaryotic cells: GFP-specific αReps are clearly sequestrated by their cognate target protein addressed to various cell compartments. These results suggest that αRep proteins with tailor-made specificity can be selected and used in living cells to track, modulate or interfere with intracellular processes

Тенденції розвитку національної інноваційної системи в Україні

Проаналізовано національну інноваційну систему України. Розглянуто галузі промисловості України за ознаками інноваційної активності та досліджено темпи зростання показників, враховуючи індекс інфляції. Встановлено, що спад темпів зростання динаміки реалізованої продукції призводить до зменшення витрат на інноваційну діяльність.Дан анализ национальной инновационной системы Украины. Рассмотрены отрасли промышленности Украины по признакам инновационной активности и исследованы темпы роста показателей, учитывая индекс инфляции. Установлено, что спад темпов роста динамики реализованной продукции приводит к уменьшению затрат на инновационную деятельность.This article analyses national innovation system of Ukraine. Examined the industry of Ukraine based on innovative activity and investigated the growth indicators, taking into account inflation-index. It is established that the slowdown in the dynamics realized production leads to a decrease in the cost of innovation

Communication of pharmacogenetic research results to HIV-infected treated patients: standpoints of professionals and patients.

International audienceThe aim of pharmacogenetic studies is to adapt therapeutic strategies to individual genetic profiles, thus maximising their efficacy and minimising the likelihood of adverse side effects. Since the advent of personalised medicine, the issue of communicating research results to participants has become increasingly important. We addressed this question in the context of HIV infection, as patients and associations are particularly concerned by research and therapeutic advances. We explored the standpoints of both research professionals and participants involved in a pharmacogenetic study conducted in a cohort of HIV-infected patients. The setting of the research protocol was followed over a 2-year period. Participants' standpoints were collected through a questionnaire and interviews were conducted with research professionals. Of 125 participants, 76% wished to receive individual results and 71% wished to receive collective results; 39% did not know when results might be expected. Communication of global research results is a principle that is generally accepted by professionals. Concerning individual feedback, the professionals felt that it was necessary if it could be of direct benefit to the participant, but they expressed doubts for situations with no recognised benefit. Our results highlight the necessity to consider this issue in greater detail. We suggest the need to anticipate the debates concerning individual feedback, to differentiate between situations and the importance of further investigations on the opportunities and modalities of communication. Finally, our work emphasised the opposite pressures between the pursuit of scientific knowledge and the therapeutic orientation of clinical trials

Imidazole propionate is increased in diabetes and associated with dietary patterns and altered microbial ecology

Microbiota-host-diet interactions contribute to the development of metabolic diseases. Imidazole propionate is a novel microbially produced metabolite from histidine, which impairs glucose metabolism. Here, we show that subjects with prediabetes and diabetes in the MetaCardis cohort from three European countries have elevated serum imidazole propionate levels. Furthermore, imidazole propionate levels were increased in subjects with low bacterial gene richness and Bacteroides 2 enterotype, which have previously been associated with obesity. The Bacteroides 2 enterotype was also associated with increased abundance of the genes involved in imidazole propionate biosynthesis from dietary histidine. Since patients and controls did not differ in their histidine dietary intake, the elevated levels of imidazole propionate in type 2 diabetes likely reflects altered microbial metabolism of histidine, rather than histidine intake per se. Thus the microbiota may contribute to type 2 diabetes by generating imidazole propionate that can modulate host inflammation and metabolism

Utilisation des

Dans les années 1980, le développement de la biologie moléculaire a permis de cloner rapidement des fragments d’anticorps recombinants sous forme de scFv (single chain Fv) fonctionnels. En raison de leur faible taille, et de leur relative facilité d’expression, les scFv présentent de nombreuses applications médicales et biotechnologiques. L’ADN codant ces anticorps peut ainsi être aisément exprimé dans des cellules de mammifères et les anticorps synthétisés ciblés vers différents compartiments cellulaires grâce à des séquences d’adressage permettant leur transport ou leur rétention dans un compartiment donné. L’anticorps recombinant peut ainsi être utilisé comme anticorps intracellulaire (intrabody) pour neutraliser, perturber ou suivre la dynamique endogène d’un antigène. Les intrabodies présentent non seulement une nouvelle voie pour la recherche fondamentale en permettant l’étude de la dynamique de protéines endogènes, mais aussi des perspectives intéressantes pour la recherche appliquée en terme d’immunisation intracellulaire en vue d’une utilisation thérapeutique