Magnetic fishing of recombinant green fluorescent proteins and tagged proteins with designed synthetic ligands

Cláudia S.M. Fernandes, Ana Sofia Pina, Íris L. Batalha, A. Cecília A. Roque

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Abstract

Biomimetic ligands have emerged to overcome disadvantages inherent in biological ligands. In particular, the Ugi reaction can generate scaffolds where molecular diversity can be introduced, allowing the synthesis and screening of ligand libraries in a high-throughput manner against a variety of biological targets. Two adsorbents bearing Ugi-based synthetic ligands, coined A4C7 and A7C1, were previously developed for the selective recovery of green fluorescent protein (GFP) and RKRKRK-tagged GFP directly from Escherichia coli crude extracts. This work describes, for the first time, the in situ synthesis of Ugi-based ligands on magnetic beads and their application in the magnetic recovery of cognate proteins.

Original languageEnglish
Pages (from-to)2907-2915
Number of pages9
JournalSeparation Science and Technology (Philadelphia)
Volume52
Issue number18
DOIs
Publication statusPublished - 12 Dec 2017

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Keywords

  • GFP purification
  • magnetic fishing
  • magnetic particles
  • synthetic ligands
  • tagged-proteins purification

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