Comparative analysis of fusion tags used to functionalize recombinant antibodies.
Binding, Competitive
Cysteine
/ metabolism
Escherichia coli
Escherichia coli Proteins
/ chemistry
Genetic Vectors
/ genetics
Green Fluorescent Proteins
/ chemistry
Oxidoreductases
/ chemistry
Protein Disulfide-Isomerases
/ chemistry
Protein Stability
Receptor, ErbB-2
/ chemistry
Recombinant Fusion Proteins
/ chemistry
Single-Domain Antibodies
/ chemistry
Antibody functionalization
Fusion tags
Nanobodies
Recombinant antibodies
Sortase
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
02 2020
02 2020
Historique:
received:
10
09
2019
revised:
20
09
2019
accepted:
25
09
2019
pubmed:
30
9
2019
medline:
23
5
2020
entrez:
30
9
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31563543
pii: S1046-5928(19)30497-8
doi: 10.1016/j.pep.2019.105505
pii:
doi:
Substances chimiques
Escherichia coli Proteins
0
Recombinant Fusion Proteins
0
Single-Domain Antibodies
0
Green Fluorescent Proteins
147336-22-9
Oxidoreductases
EC 1.-
sulfhydryl oxidase
EC 1.8.3.-
Receptor, ErbB-2
EC 2.7.10.1
Protein Disulfide-Isomerases
EC 5.3.4.1
dsbC protein, E coli
EC 5.3.4.1
Cysteine
K848JZ4886
Types de publication
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
105505Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.