Macroporous monolithic gels, cryogels, with immobilized phages from phage-display library as a new platform for fast development of affinity adsorbent capable of target capture from crude feeds.
Selected phage clones expressing a peptide with high binding affinity for recombinant human lactoferrin or von Willebrand factor (vWF) were covalently coupled to macroporous poly(dimethylacrylamide) monolithic column. Large pore size (10-100 microm) of macroporous poly(dimethylacrylamide) makes it possible to couple long (1 microm) phage particles as ligands without any risk of blocking the monolithic column. The macroporous monolithic columns were successfully used for the direct affinity capture of target proteins from particulate containing feeds like milk containing casein micelles and fat globules (1-10 microm in size) or even whole blood containing blood cells (up to 20 microm in size). The newly developed platform based on selected bacteriophages immobilized within macropores of the monolithic cryogels presents a convenient alternative to antibodies for fast and selective development of the specific adsorbent.
Noppe
Wim
W
Interdisciplinary Research Center, Katholieke Universiteit Leuven Campus Kortrijk, E. Sabbelaan 53, B-8500 Kortrijk, Belgium.
Plieva
Fatima M
FM
Vanhoorelbeke
Karen
K
Deckmyn
Hans
H
Tuncel
Mürvet
M
Tuncel
Ali
A
Galaev
Igor Yu
IY
Mattiasson
Bo
B
D001435:Bacteriophages / Q000187:drug effects / Q000502:physiology*; D001672:Biocompatible Materials / Q000737:chemistry; D001681:Biological Assay / Q000379:methods*; D001798:Blood Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D059985:Cryogels; D005353:Fibronectins / Q000737:chemistry*; D020100:Hydrogels / Q000737:chemistry*; D008422:Materials Testing; D019151:Peptide Library; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology; D016062:Porosity
D001672:Biocompatible Materials; D001798:Blood Proteins; D059985:Cryogels; D005353:Fibronectins; D020100:Hydrogels; D019151:Peptide Library; D010455:Peptides