A simple and versatile strategy for oriented immobilization of his-tagged proteins on magnetic nanoparticles

Castro-Hinojosa, Christian ; Del Sol-Fernández, Susel ; Moreno-Antolín, Eduardo ; Martín-Gracia, Beatriz ; Ovejero, Jesús G. ; Martínez de la Fuente, Jesús ; Grazú, Valeria ; Fratila, Raluca M. (Universidad de Zaragoza) ; Moros, María

Resumen: Oriented and covalent immobilization of proteins on magnetic nanoparticles (MNPs) is particularly challenging as it requires both the functionality of the protein and the colloidal stability of the MNPs to be preserved. Here, we describe a simple, straightforward, and efficient strategy for MNP functionalization with proteins using metal affinity binding. Our method involves a single-step process where MNPs are functionalized using a preformed, ready-to-use nitrilotriacetic acid-divalent metal cation (NTA-M2+) complex and polyethylene glycol (PEG) molecules. As a proof-of-concept, we demonstrate the oriented immobilization of a recombinant cadherin fragment engineered with a hexahistidine tag (6His-tag) onto the MNPs. Our developed methodology is simple and direct, enabling the oriented bioconjugation of His-tagged cadherins to MNPs while preserving protein functionality and the colloidal stability of the MNPs, and could be extended to other proteins expressing a polyhistidine tag. When compared to the traditional method where NTA is first conjugated to the MNPs and afterward free metal ions are added to form the complex, this novel strategy results in a higher functionalization efficiency while avoiding MNP aggregation. Additionally, our method allows for covalent bonding of the cadherin fragments to the MNP surface while preserving functionality, making it highly versatile. Finally, our strategy not only ensures the correct orientation of the protein fragments on the MNPs but also allows for the precise control of their density. This feature enables the selective targeting of E-cadherin-expressing cells only when MNPs are decorated with a high density of cadherin fragments.

Idioma: Inglés

DOI: 10.1021/acs.bioconjchem.3c00417

Año: 2023

Publicado en: Bioconjugate Chemistry 34, 12 (2023), 2275-2292
ISSN: 1043-1802

Factor impacto JCR: 4.0 (2023)

Categ. JCR: CHEMISTRY, ORGANIC rank: 9 / 58 = 0.155 (2023) - Q1 - T1
Categ. JCR: BIOCHEMICAL RESEARCH METHODS rank: 14 / 85 = 0.165 (2023) - Q1 - T1
Categ. JCR: CHEMISTRY, MULTIDISCIPLINARY rank: 79 / 231 = 0.342 (2023) - Q2 - T2
Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 100 / 313 = 0.319 (2023) - Q2 - T1

Factor impacto CITESCORE: 9.0 - Pharmaceutical Science (Q1) - Biotechnology (Q1) - Organic Chemistry (Q1) - Bioengineering (Q1) - Biomedical Engineering (Q1) - Pharmacology (Q1)

Factor impacto SCIMAGO: 1.085 - Bioengineering (Q1) - Biomedical Engineering (Q1) - Pharmacology (Q1) - Organic Chemistry (Q1) - Pharmaceutical Science (Q1) - Biotechnology (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/E15-20R
Financiación: info:eu-repo/grantAgreement/EC/H2020/853468/EU/Remote control of cellular signalling triggered by magnetic switching/SIROCCO
Financiación: info:eu-repo/grantAgreement/ES/MECD/FPU17-02024
Financiación: info:eu-repo/grantAgreement/ES/MICINN-AEI/PRTR-C17.I1
Financiación: info:eu-repo/grantAgreement/ES/MICINN/PGC2018-096016-B-I00
Financiación: info:eu-repo/grantAgreement/ES/MICINN/PID2021-122508NB-I00
Financiación: info:eu-repo/grantAgreement/ES/MICINN/RYC2015-17640
Financiación: info:eu-repo/grantAgreement/ES/MICINN/RYC2019-026860-I

Tipo y forma: Article (Published version)

Área (Departamento): Área Química Orgánica (Dpto. Química Orgánica)