Human Growth Hormone (HGH) Purification From CHO-Cell Culture Supernatant Utilizing Macroporous Chromatographic Media

Abstract

The human growth hormone is responsible for the growth process of human beings. In addition it plays an important role in metabolism, protein synthesis and cell proliferation. Since microsomia is one of the main consequences of hGH deficiency and occurs in one of 5000 children, the recombinant protein is the third major drug produced by biotechnology. The purification of biological active compounds such as human growth hormone from complex cell culture samples is a crucial step in proteomic research and downstream processing of biotechnological products. Traditional chromatographic methods are based on packed columns and often require several steps resulting in time consuming and costly procedures. In contrast, other chromatographic media offer some advantages: Due to their macroporous structure, monoliths and membrane adsorber based chromatographic media allow high flow rates without causing high back pressure. Thus, shear stress to fragile structures is avoided. In the media there are no long diffusion paths, mass transfer takes place through convection rather than diffusion. As a result, monolith and membrane adsorber based protein purification enables a high throughput and time effective performance. In our work we show the expression of hGH in Chinese hamster ovary (CHO) cells and its isolation from cell culture supernatant via monoliths and membrane adsorbers utilizing ion exchange and affinity chromatography.