Rapid reversed-phase high-performance liquid chromatography method for quantitation, at high pH, of the recombinant apolipoprotein A-IMilano in Escherichia coli fermentation broth.
An automated reversed-phase high-performance liquid chromatography method for quantitative determination of recombinant apolipoprotein A-IMilano (r-Apo A-IM) in E. coli fermentation broth has been developed and evaluated. The use of a unique matrix (Poros IIR/H) makes it possible to achieve rapid separation and good resolution at high pH. The r-Apo A-IM-containing fraction is well separated from other proteins allowing a reliable quantification. The automation and high sample throughput of this method makes it very useful for routine determination of r-Apo A-IM in fermentation broth and in eluates from the various purfication steps. With suitable modifications and adaptions this method is likely to be useful for similar rapid analytical determination of recombinant proteins in complex solutions.
Mulugeta
E
E
Pharmacia & Upjohn AB, Process R&D, Stockholm, Sweden.
Carnö
S
S
Becker
S
S
Hagman
A
A
Mascher
E
E
D016632:Apolipoprotein A-I / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity
D016632:Apolipoprotein A-I; D011994:Recombinant Proteins; C080631:apolipoprotein A-I Milano