Affitins for protein purification by affinity magnetic fishing
| العنوان: | Affitins for protein purification by affinity magnetic fishing |
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| المؤلفون: | A. Cecília A. Roque, Raquel Santos, Frédéric Pecorari, Aline Canani Viecinski, Barbara Mouratou, Stella Ottengy, Ghislaine Béhar, Cláudia S. M. Fernandes |
| المساهمون: | UCIBIO-REQUIMTE, Centre de Recherche en Cancérologie Nantes-Angers (CRCNA), Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM)-Hôtel-Dieu de Nantes-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Laennec-Centre National de la Recherche Scientifique (CNRS)-Faculté de Médecine d'Angers-Centre hospitalier universitaire de Nantes (CHU Nantes), This work was supported by Unidade de Ciências Biomoleculares Aplicadas, UCIBIO, financed by national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728), contracts PTDC/EBB-BIO/118317/2010, fellowships PD/BD/105871/2014 and PD/BD/105753/2014 for C. Fernandes and R. dos Santos, respectively., Bernardo, Elizabeth |
| المصدر: | Journal of Chromatography A Journal of Chromatography A, Elsevier, 2016, 1457, pp.50-8. ⟨10.1016/j.chroma.2016.06.020⟩ Journal of Chromatography A, 2016, 1457, pp.50-8. ⟨10.1016/j.chroma.2016.06.020⟩ |
| بيانات النشر: | HAL CCSD, 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, Phage display, Affitin, Archaeal Proteins, Lysozyme, Magnetic particles, [SDV.CAN]Life Sciences [q-bio]/Cancer, Ligands, 01 natural sciences, Biochemistry, Chromatography, Affinity, Antibodies, Analytical Chemistry, 03 medical and health sciences, chemistry.chemical_compound, [SDV.CAN] Life Sciences [q-bio]/Cancer, Affinity chromatography, Magnetic fishing, Protein purification, Escherichia coli, Animals, Humans, Tandem affinity purification, Chromatography, 010401 analytical chemistry, Organic Chemistry, General Medicine, Protein engineering, Affinity purification, 0104 chemical sciences, DNA-Binding Proteins, 030104 developmental biology, chemistry, Immunoglobulin G, Ribosome display, Magnets, Muramidase, Chickens, Protein Binding |
| الوصف: | International audience; Currently most economical and technological bottlenecks in protein production are placed in the downstream processes. With the aim of increasing the efficiency and reducing the associated costs, various affinity ligands have been developed. Affitins are small, yet robust and easy to produce, proteins derived from the archaeal extremophilic " 7 kDa DNA-binding " protein family. By means of combinatorial protein engineering and ribosome display selection techniques, Affitins have shown to bind a diversity of targets. In this work, two previously developed Affitins (anti-lysozyme and anti-IgG) were immobilized onto magnetic particles to assess their potential for protein purification by magnetic fishing. The optimal lysozyme and human IgG binding conditions yielded 58 mg lysozyme/g support and 165 mg IgG/g support, respectively. The recovery of proteins was possible in high yield (≥95%) and with high purity, namely ≥95% and 81%, when recovering lysozyme from Escherichia coli supernatant and IgG from human plasma, respectively. Static binding studies indicated affinity constants of 5.0 × 10 4 M −1 and 9.3 × 10 5 M −1 for the anti-lysozyme and anti-IgG magnetic supports. This work demonstrated that Affitins, which can be virtually evolved for any protein of interest, can be coupled onto magnetic particles creating novel affinity adsorbents for purification by magnetic fishing. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 0021-9673 1873-3778 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::85a2d44afae64ba83f9eedadfd445ff7 https://www.hal.inserm.fr/inserm-01342446/file/ArticlePecorari2016.pdf |
| حقوق: | EMBARGO |
| رقم الأكسشن: | edsair.doi.dedup.....85a2d44afae64ba83f9eedadfd445ff7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00219673 18733778 |
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