RAPID HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC QUANTIFICATION OF RECOMBINANT HUMAN ANTITHROMBIN-III DURING PRODUCTION AND PURIFICATION

For monitoring of recombinant human antithrombin III during cell cultu re processes and subsequent purification steps a rapid method for quan titative determination was developed. The need for the introduction of this rapid method came from the limited availability of a quantitativ e enzyme-linked immunosorbent assay (ELISA) and the very time-consumin g ELISA procedure. The developed method is based on reversed-phase hig h-performance liquid chromatography using a C-4 column. The separation by gradient elution using water and acetonitrile takes less than 20 m in even when complex samples, such as serum containing cell culture sa mples, have to be analyzed. Automation and a high sample throughput ar e possible with this reliable method. If necessary, insulin, transferr in and albumin can also be quantified with minor changes of the elutio n profile.