My watch list
my.chemeurope.com  
Login  

Effects of Bed Compression on Protein Separation on Gel Filtration Chromatography at Bench and Pilot Scale

Abstract BACKGROUND

Poorly packed chromatography columns are known to reduce drastically the column efficiency and produce broader peaks. Controlled bed compression has been suggested to be a useful approach for solving this problem. Here the relationship between column efficiency and resolution of protein separation are examined when preparative chromatography media were compressed using mechanical and hydrodynamic methods. Sepharose CL‐6B, an agarose based size exclusion media was examined at bench and pilot scale. The asymmetry and height equivalent of a theoretical plate (HETP) was determined by using 2% v/v acetone, whereas the void volume and intraparticle porosity (εp) were estimated by using blue dextran. A protein mixture of ovalbumin (chicken), bovine serum albumin (BSA) and ɣ′‐ globulin (bovine) with molecular weights of 44, 67 and 158 kDa, respectively, were used as a “model” separation challenge.

RESULTS

Mechanical compression achieved a reduction in plate height for the column with a concomitant improvement in asymmetry. Furthermore, the theoretical plate height decreased significantly with mechanical compression resulting in a 40% improvement in purity compared to uncompressed columns at the most extreme conditions of compression used.

CONCLUSION

The results suggest that the mechanical bed compression of Sepharose CL‐6B can be used to improve the resolution of protein separation.

Authors:   Darryl Y. C. Kong, Spyridon Gerontas, Ross A. McCluckie, Martin Mewies, David Gruber, Nigel J. Titchener‐Hooker
Journal:   Journal of Chemical Technology & Biotechnology
Year:   2017
Pages:   n/a
DOI:   10.1002/jctb.5411
Publication date:   11-Aug-2017
Facts, background information, dossiers
More about Wiley
Your browser is not current. Microsoft Internet Explorer 6.0 does not support some functions on Chemie.DE