Paper Chromatography of Protein Mixtures and Blood Plasmas

Summary

A technic for the two dimensional chromatography of blood plasma and protein mixtures on filter paper is described. Separations of protein constituents are greatly facilitated by the addition of surface active substances such as the “Tweens” or “Spans,” etc. The technic at present adopted is to add “Tween 85” or “Tween 81” and, using hemin as protein marker, to employ M/10 sucrose solution in the first dimension and M/10 sodium potassium tartrate solution in the second dimension. Differences between the protein patterns of chromatograms of human, rat and guinea pig plasma are noted. With protein mixtures, associations of proteins may occur that may be detected by chromatography. The sera of hyperimmunized horses (as against diphtheria toxin) yield chromatograms which differ from, and exhibit fractions not present in, chromatograms of normal horse sera.

Grateful acknowledgment is made to the National Cancer Institute of Canada for a grant in aid of this work, to the Department of Microbiology and Hygiene of the University of Montreal for gifts of normal and immune sera, to Dr. J. Gross for the protein fractions obtained from the Cutter Laboratories, Berkeley, Calif., to the Atlas Powder Co. (per G. F. Sterne & Son, Brantford, Ont.), for gifts of “Spans,” “Tweens,” “NNO,” and “BRIJ-30,” to the Canadian Industries, Ltd., Montreal, for gifts of “Elvanols,” and to Mrs. Mary Hersey for her able assistance in this work.