Retroviral particles are effectively purified on an affinity matrix containing peptides selected by phage‐display. Issue 12 (26th September 2016)

• Record Type: Journal Article
• Authors: Fernandes, Cláudia S. M.; Barbosa, Inês; Castro, Rute; Pina, Ana Sofia; Coroadinha, Ana Sofia; Barbas, Ana; Roque, A. Cecília A.
• Is Part Of: Biotechnology journal. Volume 11:Issue 12(2016)
• Page Start: 1513
• Page End: 1524
• Publication Date: 2016-09-26
• Subjects: Affinity ligands -- Peptides -- Phage display -- Virus‐like particles -- VLP purification; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201600025
• Languages: English
• ISSNs: 1860-6768

Abstract

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. Abstract : Existing viral particle purification technologies cannot distinguish between infectious and non‐infectious retroviral particles that consequently impairs the enrichment of infectious viral particles in the final purified solution. Phage display rendered a peptide which showed high differentiation between VLP‐AMPHO and VLP null. The peptide was used to functionalize cross‐linked agarose and magnetic particles. A selective capture and mild recovery of murine leukemia virus‐like particles was possible. Therefore, the results obtained can be employed in gene therapy applications.