Approaches to Bioresponse-Linked Instrumental Analysis: On-Line Coupling of Liquid Chromatography to Biological Assays

Abstract

During the recent years on-line coupling of high performance liquid chromatography (HPLC) with bioassays, has evolved into a rapid, selective and above all a highly sensitive methodology, which is characterized by a broad range of interesting applications. In contrast to conventional microtiter type assays, the combination of separation power offered by LC and the high selectivity and sensitivity featured by bioassays enables the separate quantification of cross-reactive compounds. Moreover, on-line coupling of HPLC and bioassays overcomes both tedious fraction collection and manual operations, which are required to prepare fractions for batch biological assays, thus increasing the speed of analysis dramatically. This overview presents different strategies to implement biomolecular affinity interactions in on-line biochemical detection (BCD) techniques. Initially, on-line continuous-flow BCD-systems were primarily focussed on low molecular mass targets. Recently, however, we developed BCD strategies suitable for detecting high molecular mass ligands (e.g., proteins) as well, using either labeled receptors or labeled antibodies.