Nanotech 2010

Bacteriophages and cryogels: A new efficient tool in Bioseparation

W. Noppe, I.Y. Galaev, B. Mattiasson, H. Deckmyn
Katholieke Universiteit Leuven Campus Kortrijk, BE

Keywords: bacteriophages, cryogel, affinity chromatography

Abstract:

In the field of affinity chromatography, the search for new ligands is of great interest, where apart from the use of antibodies as affinity ligands, also synthetic ligands were introduced. Special attention was focused on peptide ligands. Peptides also can be selected by screening peptide displaying phage libraries (biopanning) towards a target. New protein production technologies have also an influence on down-stream processing techniques as crude feeds impose higher demands on the chromatographic matrices. New matrices and techniques are developed to deal with this challenge, such as supermacroporous cryogel monoliths. Due to the macropores particulate feeds can flow through the cryogel without any restriction. We here used, instead of isolated peptides, the bacteriophages expressing the selected peptide on an envelope protein, directly as affinity ligands. This has several advantages when compared to a synthetic peptide: (i) once the phage is selected, it can be amplified in large quantities at low cost and (ii) by using bacteriophages a very specific ligand is available combining the effects of “affinity” and “avidity” for the optimal capture of target. We furthermore attached the bacteriophages as affinity ligands to a cryogel monolith column and provided “proof of principle” for this new technique in bio-separation.
 
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