title: Biobased monoliths for adenovirus purification.
abstract: Adenoviruses are important platforms for vaccine development and vectors for gene therapy, increasing the demand for high titers of purified viral preparations. Monoliths are macroporous supports regarded as ideal for the purification of macromolecular complexes, including viral particles. Although common monoliths are based on synthetic polymers as methacrylates, we explored the potential of biopolymers processed by clean technologies to produce monoliths for adenovirus purification. Such an approach enables the development of disposable and biodegradable matrices for bioprocessing. A total of 20 monoliths were produced from different biopolymers (chitosan, agarose, and dextran), employing two distinct temperatures during the freezing process (-20 °C and -80 °C). The morphological and physical properties of the structures were thoroughly characterized. The monoliths presenting higher robustness and permeability rates were further analyzed for the nonspecific binding of Adenovirus serotype 5 (Ad5) preparations. The matrices presenting lower nonspecific Ad5 binding were further functionalized with quaternary amine anion-exchange ligand glycidyltrimethylammonium chloride hydrochloride by two distinct methods, and their performance toward Ad5 purification was assessed. The monolith composed of chitosan and poly(vinyl) alcohol (50:50) prepared at -80 °C allowed 100% recovery of Ad5 particles bound to the support. This is the first report of the successful purification of adenovirus using monoliths obtained from biopolymers processed by clean technologies.
authors.affiliation: †UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Cláudia S M
authors.initials: CS
authors.lastname: Fernandes
authors.affiliation: †UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Bianca
authors.initials: B
authors.lastname: Gonçalves
authors.affiliation: †UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.|§LAQV, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Margarida
authors.initials: M
authors.lastname: Sousa
authors.affiliation: ‡Instituto de Biologia Experimental Tecnológica, Avenida da República, Quinta do Marquês, Edificio IBET/ITQB, Estação Agronómica Nacional, 2780-157 Oeiras, Portugal.
authors.forename: Duarte L
authors.initials: DL
authors.lastname: Martins
authors.affiliation: †UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.|§LAQV, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Telma
authors.initials: T
authors.lastname: Barroso
authors.affiliation: †UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Ana Sofia
authors.initials: AS
authors.lastname: Pina
authors.affiliation: ‡Instituto de Biologia Experimental Tecnológica, Avenida da República, Quinta do Marquês, Edificio IBET/ITQB, Estação Agronómica Nacional, 2780-157 Oeiras, Portugal.
authors.forename: Cristina
authors.initials: C
authors.lastname: Peixoto
authors.affiliation: §LAQV, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Aguiar-Ricardo
authors.affiliation: †UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: A Cecília A
authors.initials: AC
authors.lastname: Roque
mesh_terms: D000256:Adenoviridae / Q000737:chemistry* / Q000302:isolation & purification; D000327:Adsorption; D001704:Biopolymers / Q000737:chemistry*
chemical_list: D001704:Biopolymers
keywords: biopolymers; clean processes; cryogels; monoliths; plasma processing; virus purification