Affitins as robust tailored reagents for affinity chromatography purification of antibodies and non-immunoglobulin proteins
| العنوان: | Affitins as robust tailored reagents for affinity chromatography purification of antibodies and non-immunoglobulin proteins |
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| المؤلفون: | Barbara Mouratou, Frédéric Pecorari, Ghislaine Béhar, Axelle Renodon-Cornière |
| المساهمون: | 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 the Institut Pasteur (Paris, France) & Bio-Rad Laboratories, and 'La Région des Pays de la Loire' through funding DI 2006-42 and 2007-6174, respectively., Bernardo, Elizabeth |
| المصدر: | Journal of Chromatography A Journal of Chromatography A, Elsevier, 2016, 1441, pp.44-51. ⟨10.1016/j.chroma.2016.02.068⟩ Journal of Chromatography A, 2016, 1441, pp.44-51. ⟨10.1016/j.chroma.2016.02.068⟩ |
| بيانات النشر: | HAL CCSD, 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, Strep-tag, Affitin, Archaeal Proteins, [SDV.CAN]Life Sciences [q-bio]/Cancer, medicine.disease_cause, Ligands, Affinity chromatography, 01 natural sciences, Biochemistry, Chromatography, Affinity, Analytical Chemistry, Matrix (chemical analysis), 03 medical and health sciences, chemistry.chemical_compound, Bacterial Proteins, [SDV.CAN] Life Sciences [q-bio]/Cancer, medicine, Immunoglobulin, Animals, Humans, Staphylococcal Protein A, Escherichia coli, Antibody, Chromatography, biology, Ligand, Escherichia coli Proteins, 010401 analytical chemistry, Organic Chemistry, Sso7d, General Medicine, 0104 chemical sciences, 030104 developmental biology, Immobilized Proteins, chemistry, Sac7d, Immunoglobulin G, biology.protein, Agarose, Indicators and Reagents, Muramidase, Protein A, Bacterial Outer Membrane Proteins |
| الوصف: | International audience; Affinity chromatography is a convenient way of purifying proteins, as a high degree of purity can be reached in one step. The use of tags has greatly contributed to the popularity of this technique. However, the addition of tags may not be desirable or possible for the production of biopharmaceuticals. There is thus a need for tailored artificial affinity ligands. We have developed the use of archaeal extremophilic proteins as scaffolds to generate affinity proteins (Affitins). Here, we explored the potential of Affitins as ligand to design affinity columns. Affitins specific for human immunoglobulin G (hIgG), bacterial PulD protein, and chicken egg lysozyme were immobilized on an agarose matrix. The columns obtained were functional and highly selective for their cognate target, even in the presence of exogenous proteins as found in cell culture media, ascites and bacterial lysates, which result in a high degree of purity (∼95%) and recovery (∼100%) in a single step. Anti-hIgG Affitin columns withstand repetitive cycles of purification and cleaning-in-place treatments with 0.25 M NaOH as well as Protein A does. High levels of Affitin productions in Escherichia coli makes it possible to produce these affinity columns at low cost. Our results validate Affitins as a new class of tailored ligands for the affinity chromatography purification of potentially any proteins of interest including biopharmaceuticals. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 0021-9673 1873-3778 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::637f535bc1d3905f5a58f81ee31f45c2 https://www.hal.inserm.fr/inserm-01292506 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....637f535bc1d3905f5a58f81ee31f45c2 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00219673 18733778 |
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