Fiber optic multi-channel protein detector for use in preparative continuous annular chromatography

Abstract

Continuous annular chromatography is an effective method in the separation of preparative scale quantities of biological compounds including proteins where established batch chromatography borders on it. The need for identification or quantification of proteins triggered the development of respective detection units. Here, we describe two types of optical multi-channel detectors. The first is a fiber optic multi-channel detector suitable for the separation of aqueous protein solutions. The second is a technically improved (circular optic) device suitable for application in multi-channel detection. Specifically, UV-absorption measurements of proteins at 280 nm were carried out using newly designed fiber optic detectors having eight and 16 channels. Calibration plots were established for a series of stock solutions of known concentrations of proteins. Mathematical functions were derived from these calibration data to simulate the response of the detector. Limits of detection and the ranges of validity of the fit functions were determined. The 16-channel detector has a theoretical limit of 24 detection that is equivalent to absorbance changes of 10⁻⁴ units.