ANALYTICAL METHOD OPTIMIZATION FOR PROTEIN DETERMINATION BY FAST HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A method based on fast HPLC allowing the rapid and efficient determina tion of proteins in mammalian cell cultures is proposed, involving the use of two chromatographic modes, RP-HPLC and high-performance liquid affinity chromatography. These two sequential chromatographic analyse s separate the proteins most often used as supplements in serum-free m edia such as bovine serum albumin, insulin and transferrin, and also m onoclonal antibodies secreted by hybridoma in cultures. Rapidity, reli ability and flexibility are the main characteristics of the method. Th e monitoring of proteins in the course of a discontinuous hybridoma cu lture is presented.