Počet záznamů: 1
Tris-(Nitrilotriacetic Acid)-Decorated Polymer Conjugates as Tools for Immobilization and Visualization of His-Tagged Proteins
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SYSNO ASEP 0519252 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Tris-(Nitrilotriacetic Acid)-Decorated Polymer Conjugates as Tools for Immobilization and Visualization of His-Tagged Proteins Tvůrce(i) 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, ORCIDČíslo článku 1011 Zdroj.dok. Catalysts. - : MDPI
Roč. 9, č. 12 (2019)Poč.str. 11 s. Jazyk dok. eng - angličtina Země vyd. CH - Švýcarsko Klíč. slova antibody mimetics ; protein immobilization ; His-tag ; tris-nitrilotriacetic acid (tris-NTA) ; iBodies ; biocompatible polymers ; N-(2-hydroxypropyl) methacrylamide (HPMA) Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology Vědní obor RIV – spolupráce Ústav makromolekulární chemie - Makromolekulární chemie CEP EF16_019/0000729 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy FV10490 GA MPO - Ministerstvo průmyslu a obchodu FV10370 GA MPO - Ministerstvo průmyslu a obchodu GA19-10280S GA ČR - Grantová agentura ČR LQ1604 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy ED1.1.00/02.0109 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Způsob publikování Open access Institucionální podpora UOCHB-X - RVO:61388963 ; UMCH-V - RVO:61389013 UT WOS 000507336600040 EID SCOPUS 85078660535 DOI 10.3390/catal9121011 Anotace 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. Pracoviště Ústav organické chemie a biochemie Kontakt asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Rok sběru 2020 Elektronická adresa https://www.mdpi.com/2073-4344/9/12/1011
Počet záznamů: 1