Retroviral particles are effectively purified on an affinity matrix containing peptides selected by phage-display

Cláudia S. M. Fernandes, Inês Barbosa, Rute Castro, Ana Sofia Fidalgo Pombo Mendes Pina, Ana Sofia Coroadinha, Ana Barbas, A Cecília A Roque

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1513-1524
Number of pages12
JournalBiotechnology Journal
Volume11
Issue number12
DOIs
Publication statusPublished - 1 Dec 2016

Keywords

  • Affinity ligands
  • Peptides
  • Phage display
  • Virus-like particles
  • VLP purification

Fingerprint Dive into the research topics of 'Retroviral particles are effectively purified on an affinity matrix containing peptides selected by phage-display'. Together they form a unique fingerprint.

  • Cite this