High throughput screening of mixed-mode sorbents and optimisation using pre-packed lab-scale columns for the purification of the recombinant allergen rBet v 1a

摘要

Mixed-mode chromatography was investigated for the purification of the recombinant allergen rBet v la expressed in Escherichia coli (E. coli) and used as an active principle for specific immunotherapy (SIT) treatment against birch pollen allergy. The screening of micro-volumes of three mixed-mode sorbents established that rBet v la could be captured without any pre-treatment of the crude feedstock on HEA or PPA HyperCel sorbents equilibrated in "physiological-like" conditions. On a mini-column pre-packed with PPA HyperCel sorbent, rBet v la was recovered at pH 4, partially separated from a major methionine Bet v I contaminant and purified approximately 9-fold in a single step (85% purity). (C) 2009 Elsevier B.V. All rights reserved.