Spot-line chromatography: a technique for studying interactions between proteins and dyes

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Abstract
Binding between proteins and other molecules can be studied by allowing a spot of protein to overtake a line of a second component in a suitable supporting medium. Binding may be shown by a distortion or loop in the line. This effect has been used to screen a number of dyes for interaction with bovine serum albumin and ovalbumin. It has been shown that under ideal conditions the area of the loop formed by a given protein in a given line of dye should be proportional to the total weight of active protein in the spot. This has been verified as being approximately true for the bovine serum albumin-Biebrich scarlet system, but with one important reservation. As bovine serum albumin molecules move across filter paper, their binding power for Biebrich scarlet falls to about 10% of the initial value. This seems to be due to adsorptive interaction between the protein and the paper.