Direct Combination of a High Performance Liquid Chromatograph and a Circular Dichroism Spectrometer for Separation and Structural Analysis of Proteins

Abstract

Basic studies of the combined system of a high performance liquid chromatograph (HPLC) and a circular dichroism (CD) spectrometer for separation and analysis of proteins are described. The HPLC-CD measurement of standard protein mixture was easily carried out by using a micro flow-cell device with a beam condenser and with a thin cell of a 1 mm-optical path. The effluent was firstly monitored at 280 nm by using an UV detector and subsequently monitored at 220 nm by using a CD spectrometer. The CD spectrum at each chromatographic peak by CD was measured in the wavelength region of 250–195 nm by a stopped flow method.