Characterization of pituitary and peptide hormones by electrophoresis in starch gel.

In 1957, Poulik (I) described a modification of Smithies’ (2) method of starch gel electrophoresis incorporating a discontinuous buffer system. In his original presentation, Poulik described a peculiar and as yet not fully explained phenomenon of an advancing borate “front” which seemed to be a major factor responsible for the high degree of separation and resolution. In 1960, Ferguson and Wallace (3, 4), using a slightly different buffer system, substituting lithium hydroxide for the sodium hydroxide of Poulik’s buffer system, and using thin 3-mm horizontal sheets of starch gel with high voltage under refrigeration, described a method of remarkably high resolution for the separation of pituitary peptides. Further evidence of the high degree of resolution of this method was the observation of Morris’ that “pure” As hemoglobin, which produced a single benzidine-positive component by the more usual starch gel electrophoresis, was separated by the Ferguson and Wallace method into four distinct benzidine-positive components and two other benzidinenegative bands. Our main interest in this method has been to follow the purification of pituitary hormones, and it has been most useful for this purpose. This report deals primarily with a description of the method and the characteristics of a variety of peptide hormone preparations thus far examined.