دورية أكاديمية
Comparative analysis of fusion tags used to functionalize recombinant antibodies.
| العنوان: | Comparative analysis of fusion tags used to functionalize recombinant antibodies. |
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| المؤلفون: | Veggiani G; Lab of Environmental and Life Sciences, University of Nova Gorica, Vipavska cesta 13, 5000, Rožna Dolina, Nova Gorica, Slovenia., Giabbai B; Structural Biology Lab, Elettra Sincrotrone Trieste S.C.p.A., 34149, Basovizza, Trieste, Italy., Semrau MS; Structural Biology Lab, Elettra Sincrotrone Trieste S.C.p.A., 34149, Basovizza, Trieste, Italy., Medagli B; Department of Chemical and Pharmaceutical Sciences, University of Trieste, Via L. Giorgieri 1, 34127, Trieste, Italy., Riccio V; Structural Biology Lab, Elettra Sincrotrone Trieste S.C.p.A., 34149, Basovizza, Trieste, Italy., Bajc G; Department of Biology, Biotechnical Faculty, University of Ljubljana, Večna pot 111, 1000, Ljubljana, Slovenia., Storici P; Structural Biology Lab, Elettra Sincrotrone Trieste S.C.p.A., 34149, Basovizza, Trieste, Italy., de Marco A; Lab of Environmental and Life Sciences, University of Nova Gorica, Vipavska cesta 13, 5000, Rožna Dolina, Nova Gorica, Slovenia. Electronic address: ario.demarco@ung.si. |
| المصدر: | Protein expression and purification [Protein Expr Purif] 2020 Feb; Vol. 166, pp. 105505. Date of Electronic Publication: 2019 Sep 26. |
| نوع المنشور: | Comparative Study; Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 9101496 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0279 (Electronic) Linking ISSN: 10465928 NLM ISO Abbreviation: Protein Expr Purif Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Orlando, FL : Academic Press Original Publication: San Diego : Academic Press, c1990- |
| مواضيع طبية MeSH: | Recombinant Fusion Proteins/*genetics , Single-Domain Antibodies/*genetics, Binding, Competitive ; Cysteine/metabolism ; Escherichia coli ; Escherichia coli Proteins/chemistry ; Escherichia coli Proteins/genetics ; Genetic Vectors/genetics ; Green Fluorescent Proteins/chemistry ; Green Fluorescent Proteins/genetics ; Oxidoreductases/chemistry ; Oxidoreductases/genetics ; Protein Disulfide-Isomerases/chemistry ; Protein Disulfide-Isomerases/genetics ; Protein Stability ; Receptor, ErbB-2/chemistry ; Receptor, ErbB-2/genetics ; Recombinant Fusion Proteins/chemistry ; Single-Domain Antibodies/chemistry |
| مستخلص: | Recombinant antibodies can be expressed as fusion constructs in combination with tags which simplify their engineering into reliable and homogeneous immunoreagents by allowing site-specific, 1:1 functionalization. Several tags and corresponding reagents for recombinant protein derivatization have been proposed but benchmarking surveys for the evaluation of their effect on the characteristics of recombinant antibodies have not been reported. In this work we evaluated the impact on expression yields, shelf-stability, thermostability and binding affinity of a set of C-terminal tags fused to the same anti-Her2 nanobody. Furthermore, we assessed the efficiency of the derivatization process. The constructs always bore a 6xHis tag plus either the controls (EGFP and C-tag) or CLIP, HALO, AviTag, the LEPTG sequence recognized by Sortase A (Sortase tag), or a free cysteine. The advantages and drawbacks of the different systems were analyzed and discussed. (Copyright © 2019 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Antibody functionalization; Fusion tags; Nanobodies; Recombinant antibodies; Sortase |
| المشرفين على المادة: | 0 (Escherichia coli Proteins) 0 (Recombinant Fusion Proteins) 0 (Single-Domain Antibodies) 147336-22-9 (Green Fluorescent Proteins) EC 1.- (Oxidoreductases) EC 1.8.3.- (sulfhydryl oxidase) EC 2.7.10.1 (Receptor, ErbB-2) EC 5.3.4.1 (Protein Disulfide-Isomerases) EC 5.3.4.1 (dsbC protein, E coli) K848JZ4886 (Cysteine) |
| تواريخ الأحداث: | Date Created: 20190930 Date Completed: 20200522 Latest Revision: 20200522 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.pep.2019.105505 |
| PMID: | 31563543 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1096-0279 |
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| DOI: | 10.1016/j.pep.2019.105505 |