title: Retroviral particles are effectively purified on an affinity matrix containing peptides selected by phage-display.
abstract: Retroviral particles are expensive to manufacture, mostly due to the downstream processing steps which result in low recoveries (≈30%) and concentration factors. In this work, a dodecapeptide phage-display library was panned against retrovirus like particles expressing the envelope protein Ampho4070A (VLPs-AMPHO) and VLPs without the target protein, used as a negative control (VLPs). A depletion/selection panning protocol was successfully used to deal with the structural complexity of the target, and a total of three distinct peptide sequences displaying preferential binding towards VLPs-AMPHO were found. Peptide 3 (CAAALAKPHTENHLLT), which appeared as one lead candidate, was synthesized and immobilized onto two purification matrices, cross-linked agarose and magnetic particles. The matrices selectively bound VLPs-AMPHO and in both cases recovery yields higher than 90% were obtained when employing mild elution conditions, while maintaining viral particle morphology and size.
authors: UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnológia, Universidade Nova de Lisboa, Caparica, Portugal.
authors: Cláudia S M
authors: CS
authors: Fernandes
authors: iBET, Instituto de Biologia Experimental e Tecnológia, Oeiras, Portugal.
authors: Inês
authors: I
authors: Barbosa
authors: iBET, Instituto de Biologia Experimental e Tecnológia, Oeiras, Portugal.|Instituto de Tecnológia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors: Rute
authors: R
authors: Castro
authors: UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnológia, Universidade Nova de Lisboa, Caparica, Portugal.
authors: Ana Sofia
authors: AS
authors: Pina
authors: iBET, Instituto de Biologia Experimental e Tecnológia, Oeiras, Portugal.|Instituto de Tecnológia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors: Ana Sofia
authors: AS
authors: Coroadinha
authors: iBET, Instituto de Biologia Experimental e Tecnológia, Oeiras, Portugal.|Bayer Portugal, S.A., Carnaxide, Portugal.
authors: Ana
authors: A
authors: Barbas
authors: UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnológia, Universidade Nova de Lisboa, Caparica, Portugal.
authors: A Cecília A
authors: AC
authors: Roque
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D012190:Retroviridae; D050611:Sodium-Phosphate Cotransporter Proteins, Type III / Q000737:chemistry / Q000378:metabolism; D014771:Virion / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D019151:Peptide Library; D010455:Peptides; D050611:Sodium-Phosphate Cotransporter Proteins, Type III
keywords: Affinity ligands; Peptides; Phage display; VLP purification; Virus-like particles