Patent ID: 8349615
Filing Date: 2013-01-08
Classification: B01D,G01N

Abstract:
1. A method of finding suitable parameters for chromatographic processes for separating biomolecules, wherein tests comprising a sequence of equilibration, charging, washing and elution steps are carried out on a small scale, with variation of individual parameters, and from the results of these tests conclusions are drawn as to optimum parameters, characterised in that the sequence of equilibration, charging, washing and elution steps is carried out by the partial batch method (steps with and without suspension of the chromatography material), wherein at least the charging step comprises suspending the chromatography material and at least during an elution step there is no suspension of the chromatography material, wherein (a) a suspension of a chromatography material is placed in a plurality of wells in a multi-well filter plate, (b) a moist gel bed is produced in the wells containing the suspension, by removing the supernatant above the chromatography material by centrifugation or by the application of a pressure differential, (c) in order to equilibrate the chromatography material contained in the wells an equilibrating buffer solution is added to the moist gel bed and the gel bed is suspended (equilibration step), (d) a moist gel bed is produced according to step (b), (e) optionally steps (c) and (d) are repeated several times, in particular once, twice or three times, (f) in order to charge the chromatography material a test solution containing at least one biomolecule is added to the gel bed, and the gel bed is suspended (charging step), (g) a moist gel bed is produced according to step (b), (h) in order to wash the chromatography material a washing solution is added to the gel bed, the gel bed optionally being suspended (washing step), (i) a moist gel bed is produced according to step (b), (k) optionally steps (h) and (i) are repeated several times, particularly once, twice or three times, while the washing solutions used may have the same or different compositions and optionally the gel bed is suspended or is not suspended, (l) for eluting the biomolecule, an eluting solution is added to the gel bed without causing suspension of the gel bed (elution step), (m) according to step (b) a moist gel bed is produced and the eluate is caught, (n) optionally steps (l) and (m) are repeated several times, particularly once, twice or three times, while the eluant solutions used may have the same or different compositions, and (o) the eluates collected are analysed.