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.
J Chromatogr B Analyt Technol Biomed Life Sci
; 877(24): 2420-7, 2009 Aug 15.
Article
in En
| MEDLINE
| ID: mdl-19345650
ABSTRACT
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).
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Pollen
/
Resins, Synthetic
/
Allergens
/
Chromatography, Liquid
/
Betula
Type of study:
Diagnostic_studies
/
Evaluation_studies
/
Screening_studies
Limits:
Humans
Language:
En
Journal:
J Chromatogr B Analyt Technol Biomed Life Sci
Journal subject:
ENGENHARIA BIOMEDICA
Year:
2009
Document type:
Article