Application of image analysis techniques to two-dimensional crossed immunoelectrophoresis study

Two-dimensional crossed immunoelectrophoresis (2D-CIEP) is a technique widely used for studying composition of protein mixtures in biological and clinical studies. A low cost image analysis system with the use of an optical flatbed scanner and a IBM-compatible PC was set up in this work for capturing 2D-CIEP patterns. A computer package CIEPEASY was developed for modification and analysis of the image acquired to determine various peak parameters such as the migration distance, peak height and peak area of the constituting components for both qualitative and quantitative studies. In this approach, more composition information of the standard and sample gel patterns could be extracted from the proposed image analysis system. The time required for data collection and interpretation of 2D-CIEP images was shortened significantly and the results obtained have a higher accuracy than those obtained by using conventional methods. Moreover, a linear relationship between the peak area and the amount of antigen present in a sample was confirmed accurately and reported for the first time in the literature.