Single step purification of a series of wheat recombinant proteins with expanded bed absorption chromatography.
Expanded bed absorption chromatography (EBA) was used to improve and simplify the purification of several wheat recombinant proteins. Binding and elution conditions were set to allow the purification of the over expressed protein in a single step. In comparison with our previous multi step protocol, same purity was obtained while EBA required less time (one day instead of five) and gave a higher yield (63% instead of 10%). This new procedure was then used for the successful purification of five other wheat ns-LTP. Despite their important polymorphism (identity from 44 to 97 %-pHi from 8 to 10), the EBA protocol allowed their purification in a single step.
de Lamotte
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UMR Polymorphismes d'Intérêts Agronomiques-1096, INRA/CIRAD/ENSA-M, 2 Place Viala, 34060 Montpellier, Cedex 01, France. lamotte@ensam.inra.fr
D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D008565:Membrane Proteins / Q000302:isolation & purification; D049294:Phospholipid Transfer Proteins / Q000302:isolation & purification; D010843:Pichia / Q000378:metabolism; D010940:Plant Proteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D014908:Triticum / Q000378:metabolism*
D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins