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A robust affinity chromatography system based on ceramic monoliths coated with poly(amino acid)‐based polymeric constructs
Author(s) -
Santiago Javier Sánchez,
Cerro Ramón L,
Scholz Carmen
Publication year - 2021
Publication title -
polymer international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.592
H-Index - 105
eISSN - 1097-0126
pISSN - 0959-8103
DOI - 10.1002/pi.6142
Subject(s) - monolith , elution , acrylic acid , adsorption , polymerization , monolithic hplc column , chromatography , materials science , ceramic , asparagine , chemical engineering , copolymer , protein adsorption , polymer chemistry , chemistry , polymer , amino acid , organic chemistry , high performance liquid chromatography , catalysis , biochemistry , engineering
Traditional chromatographic separation systems are disadvantaged by low flow rates, a high pressure drop across the column, low capacity and poor reusability. Searching for more efficient separation systems we introduced the use of a ceramic monolith as robust support in bioseparations. A coating consisting of l ‐asparagine as ligand, poly( l ‐lysine) as spacer arm and a commercial poly(ethylene acrylic acid) film forming copolymer network (Michem 4983‐40R) was developed as a coating for these ceramic monoliths. Poly( l ‐lysine) was synthesized by ring‐opening polymerization of ε‐trifluoroacetyl‐ l ‐lysine N ‐carboxyanhydride and coupled to a commercial film‐forming poly(ethylene acrylic acid) network. This construct was then ‘decorated’ with l ‐asparagine via the terminal amino functional groups of poly(L‐lysine) and coated onto the ceramic monolith to selectively bind l ‐asparaginase. Adsorption/elution experiments showed reversible binding between l ‐asparagine and l ‐asparaginase, and the subsequent release of l ‐asparaginase, and between 83% and 94% of the active enzyme was recovered by elution with d ‐asparagine and NaCl solutions. The functional activity of the eluted l ‐asparaginase was verified by a Nessler's assay. While traditional separation processes (adsorption and elution) using gel bead packings take many hours, the ceramic monolith system achieves the same of level of separation in about 1 h. This new system served as a proof of concept for its application in protein separation and purification. This work paves the way to a better understanding of the use of ceramic monoliths as stationary phase coated with a stable polymer construct for more robust and efficient supports in affinity chromatography. © 2020 Society of Industrial Chemistry