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

作者：Brochier Virginie Brenac^*; Chabre Henri; Lautrette Aurelie; Ravault Vincent; Couret Marie Noelle; Didierlaurent Alain; Moingeon Philippe

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2009, 877(24): 2420-2427.

DOI：10.1016/j.jchromb.2009.03.020

• Recombinant allergen
• rBet v 1 a
• High-throughput screening
• Mixed-mode sorbents
• HEA HyperCel
• PPA HyperCel
• MEP HyperCel
• Pre-packed columns

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).

• 出版日期2009-8-15