Paper Chromatography of Protein Mixtures and Blood Plasmas

Abstract

Summary

Further details are given concerning the movements of proteins on filter paper using M/10 solutions of sucrose solution and sodium potassium tartrate as developing solvents in the first and second dimension, respectively. It is shown that mixtures of proteins, such as serum albumin and cytochrome c, may be separated from each other on paper, since the former moves in the first dimension, whilst the latter does not and both proteins move in the second dimension. The amount of cytochrome c moving in the second dimension is proportional to the amount applied to the paper. The chromatographic patterns of purified proteins e.g. crystalline human serum albumin, α, β, and γ globulins, and fibrinogen differ from each other in a characteristic manner. The differences are more marked when the proteins are mixed with surface active agents with which they form complexes; and these complexes move at different rates on filter paper. Complexes of surface active agents with albumin move with greater ease along the first dimension than those formed with globulins; the latter tend to remain at the point of origin. Almost all the protein complexes, however, move in the second dimension. The pattern given with a blood plasma is a composite of the patterns of the individual protein fractions, the basal fractions being apparently due to globulin. The patterns of chromatograms of pathological cases (multiple myeloma, liver cirrhosis, cancer of the stomach, lupus erythematosis) differ significantly from the normal, and are often characterized by the presence of heavy basal fractions due probably to increase in the globulin content of the plasma.

The chromatograms of dried snake venoms (cobra, Russel viper, Agkistrodon piscivorus) show various distinctive features.