On-a-chip tryptic digestion of transthyretin: a step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis
| العنوان: | On-a-chip tryptic digestion of transthyretin: a step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis |
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| المؤلفون: | jeanne bataille, Antoine Pallandre, Jörg Peter Kutter, Arthur Viode, Stéphanie Descroix, Josiane P. Lafleur, François Becher, Isabelle Le Potier, Fanny Varenne, Myriam Taverna, Christian Poüs, Iago Pereiro, camille roesch, Claire Smadja |
| المساهمون: | Université Paris Saclay (COmUE), Faculté de Pharmacie, Université Paris-Sud - Paris 11 (UP11), Institut Curie [Paris], University of Copenhagen = Københavns Universitet (KU), IZON Sciences Ldt, Partenaires INRAE, Hôpital Antoine Béclère, Assistance Publique - Hôpitaux de Paris (AP-HP), Laboratoire de Chimie Physique D'Orsay (LCPO), Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre de Nanosciences et Nanotechnologies (C2N (UMR_9001)), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), fellowship programme of DIM Analytics, DIGIDIAG projects - French National Research Agency (ANR) as part of the Investissements d'Avenir programme [ANR 10-NANO 0207] |
| المصدر: | Analyst Analyst, Royal Society of Chemistry, 2018, 143 (5), pp.1077-1086. ⟨10.1039/c7an01737e⟩ |
| بيانات النشر: | HAL CCSD, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, proteolysis, Hydrochloride, diagnosis, monoliths, [SDV]Life Sciences [q-bio], Peptide, protein digestion, 01 natural sciences, Biochemistry, magnetic beads, Analytical Chemistry, 03 medical and health sciences, Hydrolysis, chemistry.chemical_compound, Electrochemistry, medicine, solvent systems, Humans, Prealbumin, Environmental Chemistry, Monolith, Spectroscopy, chemistry.chemical_classification, enzyme reactors, Amyloid Neuropathies, Familial, geography, geography.geographical_feature_category, Chromatography, biology, Chemistry, 010401 analytical chemistry, high-efficiency, Reproducibility of Results, Substrate (chemistry), nutritional and metabolic diseases, Microfluidic Analytical Techniques, mass-spectrometry, Trypsin, 0104 chemical sciences, Transthyretin, trypsin, 030104 developmental biology, biology.protein, Microreactor, medicine.drug |
| الوصف: | International audience; A microfluidic microreactor for trypsin mediated transthyretin (TTR) digestion has been developed as a step towards the elaboration of a fully integrated microdevice for the detection of a rare and disabling disease, the familial transthyretin amyloidosis (ATTR) which is related to specific TTR mutations. Therefore, an enzymatic microreactor coupled to an analytical step able to monitor the mutation of TTR on specific peptide fragments would allow an accurate monitoring of the treatment efficiency of ATTR. In this study, two types of immobilized trypsin microreactors have been investigated: a new miniaturized, microfluidic fluidized bed packed with trypsin functionalized magnetic particles (MPs), and a thiol-ene (TE) monolithbased chip. Their performances were first demonstrated with N-benzoyl-DL-arginine-4-nitroanilide hydrochloride BApNA, a low molecular weight substrate. High reaction yields (75.2%) have been reached within 0.6 min for the TE-based trypsin microreactor, while a lower yield (12.4%) was obtained for the micro-fluidized bed within a similar residence time. Transposition of the optimized conditions, developed with BApNA, to TTR digestion in the TE-based trypsin microreactor was successfully performed. We demonstrated that the TE-chip can achieve an efficient and reproducible digestion of TTR. This has been assessed by MS detection. In addition, TTR hydrolysis led to the production of a fragment of interest allowing the therapeutic follow-up of more than twenty possible ATTR mutations. High sequence coverage (90%), similar to those obtained with free trypsin, was achieved in a short time (2.4 min). Repeated experiments showed good reproducibility (RSD = 6.8%). These promising results open up the route for an innovative treatment follow-up dedicated to ATTR. |
| اللغة: | English |
| تدمد: | 0003-2654 1364-5528 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8b5559964dbb29c3d394573545fce056Test https://hal.inrae.fr/hal-02628957Test |
| حقوق: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....8b5559964dbb29c3d394573545fce056 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00032654 13645528 |
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