Expanding one-pot cell-free protein synthesis and immobilization for on-demand manufacturing of biomaterials

Benítez-Mateos, A.I. ; Llarena, I. ; Sánchez-Iglesias, A. ; López-Gallego, F. (Universidad de Zaragoza)

Resumen: Fabrication of protein-based biomaterials is an arduous and time-consuming procedure with multiple steps. In this work, we describe a portable toolkit that integrates both cell-free protein synthesis (CFPS) and protein immobilization in one pot just by mixing DNA, solid materials, and a CFPS system. We have constructed a modular set of plasmids that fuse the N-terminus of superfolded green fluorescent protein (sGFP) with different peptide tags (poly(6X)Cys, poly(6X)His, and poly(6X)Lys), which drive the immobilization of the protein on the tailored material (agarose beads with different functionalities, gold nanorods, and silica nanoparticles). This system also enables the incorporation of azide-based amino acids into the nascent protein for its selective immobilization through copper-free click reactions. Finally, this technology has been expanded to the synthesis and immobilization of enzymes and antibody-binding proteins for the fabrication of functional biomaterials. This synthetic biological platform has emerged as a versatile tool for on-demand fabrication of therapeutic, diagnostic, and sensing biomaterials.

Idioma: Inglés

DOI: 10.1021/acssynbio.7b00383

Año: 2018

Publicado en: ACS Synthetic Biology 7, 3 (2018), 875-884
ISSN: 2161-5063

Factor impacto JCR: 5.571 (2018)

Categ. JCR: BIOCHEMICAL RESEARCH METHODS rank: 6 / 78 = 0.077 (2018) - Q1 - T1

Factor impacto SCIMAGO: 2.722 - Biochemistry, Genetics and Molecular Biology (miscellaneous) (Q1) - Medicine (miscellaneous) (Q1) - Biomedical Engineering (Q1)

Financiación: info:eu-repo/grantAgreement/EUR/COST/CM1303
Financiación: info:eu-repo/grantAgreement/ES/MINECO/BIO2014-61838-EXP

Tipo y forma: Article (PostPrint)

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