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.
Mixed-mode chromatography was investigated for the purification of the recombinant allergen rBet v 1a 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 1a 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 1a was recovered at pH 4, partially separated from a major methionine Bet v 1 contaminant and purified approximately 9-fold in a single step (85% purity).
Brenac Brochier
Virginie
V
Pall Life Sciences, 48, avenue des Genottes, 95800 Cergy St Christophe, France. virginie brochier@europe.pall.com
Chabre
Henri
H
Lautrette
Aurélie
A
Ravault
Vincent
V
Couret
Marie-Noëlle
MN
Didierlaurent
Alain
A
Moingeon
Philippe
P
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D029662:Betula / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D011058:Pollen / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012117:Resins, Synthetic / Q000737:chemistry*; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology
D000485:Allergens; D011994:Recombinant Proteins; D012117:Resins, Synthetic