Immuno)affinity chromatography: a versatile tool for fast and selective purification, concentration, isolation and analysis.

摘要

Today, thanks to the availability of tailor made biomolecules with the desired biological functions, separations based upon (immuno)affinity techniques are more and more common in a large field of applications. By using the high selectivity of biomolecules (antibodies, receptors, specific proteins), this technique offers the possibility of isolating compounds from complex samples with a selectivity which cannot be achieved by other chromatographic methods. In order to succeed, however, the solid phase support for the immobilisation of the ligand of interest plays a prominent role. For this reason, numerous supports have already been introduced while research on new materials with additional advantages is continued. Here, a new solid phase support will be discussed for (immuno)affinity applications. This material demonstrates low non-specific adsorption and high ligand accessibility, which enables an enhanced selectivity and capacity. Because the material is available in large quantities and exhibits superb mechanical and physical stability, selective isolations have been performed on analytical as well as preparative scale. To demonstrate the potential of this new support, several applications will be presented. Based upon immunoaffinity, two applications for the determination of oestradiol in serum respectively vitamin B12 in fermentation broth will be presented. Regarding affinity chromatography, an enzyme reactor in which the enzyme glucose oxidase is immobilised on the new material, is made for the detection of glucose by Flow Injection Analysis and electrochemical detection. Next, to isolate, identify and test components on their xeno-oestrogenic activity, an affinity column is produced in which human oestrogen receptor is covalently coupled. Several components are screened on their biological activity and the results obtained will be presented here.