Purification of the Gamma Globulin Characteristic of Cerebrospinal Fluid

Experiments are described which have led to the standardization of a procedure for the purification of the γ c globulin characteristic of cerebrospinal fluid (CSF). The procedure involves the stepwise elution of three chromatographic fractions from a DEAE column by TRIS–HCl buffers at pH 8.5. The composition of the fractions will vary with the relative concentration of the γ c globulin in the CSF applied to the column. From an average CSF pool the first fraction, F 1 , eluted by 0.002 M TRIS–HCl will usually contain only the γ c globulin. The second fraction F 2 , eluted by 0.02 M TRIS-HCl, will contain a mixture of the γ c , γ s , and β c globulins. The third fraction F 3 , eluted by 0.08 M TRIS–HCl, will contain chiefly γ s globulin as well as traces of the γ c and β c globulins. Most of the γ c globulin from pooled CSF may then be obtained in purified form by rechromatographing the F 2 's from several runs. Almost all of the γ c globulin applied will be found in F 1 . The γ s and β c globulins will be eluted in the other two fractions.