Tris-(Nitrilotriacetic Acid)-Decorated Polymer Conjugates as Tools for Immobilization and Visualization of His-Tagged Proteins

Beranová, Jana; Knedlík, Tomáš; Šimková, Adéla; Šubr, Vladimír; Kostka, Libor; Etrych, Tomáš; Šácha, Pavel; Konvalinka, Jan

doi:https://dx.doi.org/10.3390/catal9121011

Number of the records: 1

Tris-(Nitrilotriacetic Acid)-Decorated Polymer Conjugates as Tools for Immobilization and Visualization of His-Tagged Proteins

1.

SYSNO ASEP	0519252
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Tris-(Nitrilotriacetic Acid)-Decorated Polymer Conjugates as Tools for Immobilization and Visualization of His-Tagged Proteins
Author(s)	Beranová, Jana (UOCHB-X)_ORCID Knedlík, Tomáš (UOCHB-X)_{RID, ORCID} Šimková, Adéla (UOCHB-X)_{ORCID, RID} Šubr, Vladimír (UMCH-V)_{RID, ORCID} Kostka, Libor (UMCH-V)_{RID, ORCID} Etrych, Tomáš (UMCH-V)_{RID, ORCID} Šácha, Pavel (UOCHB-X)_{RID, ORCID} Konvalinka, Jan (UOCHB-X)_{RID, ORCID}
Article number	1011
Source Title	Catalysts. - : MDPI Roč. 9, č. 12 (2019)
Number of pages	11 s.
Language	eng - English
Country	CH - Switzerland
Keywords	antibody mimetics ; protein immobilization ; His-tag ; tris-nitrilotriacetic acid (tris-NTA) ; iBodies ; biocompatible polymers ; N-(2-hydroxypropyl) methacrylamide (HPMA)
Subject RIV	CE - Biochemistry
OECD category	Biochemistry and molecular biology
Subject RIV - cooperation	Institute of Macromolecular Chemistry - Macromolecular Chemistry
R&D Projects	EF16_019/0000729 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	FV10490 GA MPO - Ministry of Industry and Trade (MPO)
	FV10370 GA MPO - Ministry of Industry and Trade (MPO)
	GA19-10280S GA ČR - Czech Science Foundation (CSF)
	LQ1604 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Method of publishing	Open access
Institutional support	UOCHB-X - RVO:61388963 ; UMCH-V - RVO:61389013
UT WOS	000507336600040
EID SCOPUS	85078660535
DOI	10.3390/catal9121011
Annotation	Recombinant proteins are commonly expressed with artificial affinity tags for purification, immobilization and characterization. The most frequently used tag, His-tag, is a sequence of consecutive histidine residues fused to the protein of interest. Specialized small molecules that bind His-tag are primarily used for purification, while antibodies are used for protein analysis. However, various issues may be encountered with the use of antibodies. Low inherent stability, the difficulty of introducing chemical modifications, and often-unreliable batch-to-batch consistency are among the limiting factors that call for better alternatives. Recently described polymer conjugates of N-(2-hydroxypropyl) methacrylamide and low-molecular-weight functional ligands, so-called iBodies, are antibody mimetics capable of replacing antibodies in biochemical applications. We tailored this system for methods utilizing His-tag by accessorizing the polymer carrier with tris-nitrilotriacetic acid targeting ligands. These anti-polyHis iBodies are additionally accessorized with fluorophores, enabling detection, and biotin ligands, enabling immobilization. Here, we characterized anti-polyHis iBodies and explored their use as antibody mimetics. We tested their stability, as well as the influence of different metal mediators and His-tag lengths on binding. With high affinity and stability, iBodies represent a new alternative for immobilization and visualization of His-tagged proteins.
Workplace	Institute of Organic Chemistry and Biochemistry
Contact	asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418
Year of Publishing	2020
Electronic address	https://www.mdpi.com/2073-4344/9/12/1011

Number of the records: 1