Isolation of Specific and Nonspecific Components from Purified Protein Derivative1,2

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Abstract
Purified protein derivative of Mycobacterium tuberculosis was separated by Sephadex G-200 into 3 fractions. These fractions were tested for sensitin activity in guinea pigs sensitized to Mycobacterium tuberculosis (homologous animals) or to Mycobacterium kansasii (heterologous animals). Fraction I contained nonspecific material. The cross reactions averaging 10.6 mm were approximately equal to the homologous reactions of 9.6 mm. This fraction contained two serologically active components, both neutral in electrophoretic mobility. Fraction II was more specific; the mean homologous reaction was 12.1 mm and the cross reaction was 4.4 mm. Immunoelectrophoresis demonstrated that fraction II contained two cathodic moving components. Fraction III was also more specific with a mean homologous reaction of 9.8 mm and a 1.8 mm cross reaction. Fraction III contained a single anodic moving component. Immunodiffusion showed that each fraction contained at least one unique element. These results suggest that purified protein derivative contains specific and nonspecific components that can be separated by a relatively simple technique.

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