Applications of ultrafast HPLC to process development of recombinant DNA-derived proteins.

The application of ultrafast HPLC to the development of recovery processes for proteins produced by recombinant DNA technology has been explored using wide-pore HPLC resins and instrumentation designed for rapid analysis. High-resolution analysis of complex samples was achieved with a total analysis time of less than 5 min from injection to injection. Fractions collected during preparative chromatography were analyzed by SDS gel electrophoresis and fast HPLC. Specific proteins in the fractions were detected and quantitated by fast HPLC providing real-time analysis for pooling. The technique was also applied to the formidable task of detecting and quantitating protein variants during the development of recovery processes. Several examples of post-translational variant detection are shown. Ultrafast HPLC is a new analytical tool that can be applied to the development of robust manufacturing processes producing therapeutic proteins essentially free of known impurities and variants.