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Bioinspired and Sustainable Chitosan Based Monoliths for Antibody Capture and Release
Publication . Barroso, Telma; Roque, Ana Cecília Afonso; Ricardo, Ana Isabel Nobre Martins Aguiar de Oliveira; DQ - Departamento de Química; CQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE); RSC - Royal Society of Chemistry
Chitosan-based monoliths activated by plasma technology induced the coupling of a robust biomimetic ligand, previously reported as an artificial Protein A, with high yields while minimizing the environmental impact of the procedure. Due to the high porosity, good mechanical and tunable physicochemical properties of the affinity chitosan-based monoliths, it is possible to achieve high binding capacities (150 ± 10 mg antibody per gram support), and to recover 90 ± 5% of the bound protein with 98% purity directly from cell-culture extracts. Therefore, the chitosan-based monoliths prepared by clean processes exhibit a remarkable performance for the one-step capture and recovery of pure antibodies or other biological molecules with biopharmaceutical relevance.
Dextran-Coated Magnetic Supports Modified with a Biomimetic Ligand for IgG Purification
Publication . Santana, Sara D. F.; Dhadge, Vijaykumar L; Roque, Ana Cecília Afonso; CQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE); DQ - Departamento de Química; ACS - American Chemical Society
Dextran-coated iron oxide magnetic particles modified with ligand 22/8, a protein A mimetic ligand, were prepared and assessed for IgG purification. Dextran was chosen as the agent to modify the surface of magnetic particles by presenting a negligible level of nonspecific adsorption. For the functionalization of the particles with the affinity ligand toward antibodies, three methods have been explored. The optimum coupling method yielded a theoretical maximum capacity for human IgG calculated as 568 ± 33 mg/g and a binding affinity constant of 7.7 × 10⁴ M⁻¹. Regeneration, recycle and reuse of particles was also highly successful for five cycles with minor loss of capacity. Moreover, this support presented specificity and effectiveness for IgG adsorption and elution at pH 11 directly from crude extracts with a final purity of 95% in the eluted fraction.
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Fundação para a Ciência e a Tecnologia
Programa de financiamento
Concurso para Projectos de I&D em todos os Domínios Científicos - 2008
Número da atribuição
PTDC/EBB-BIO/098961/2008