PURIFICATION OF THE GAMMA GLOBULIN CHARACTERISTIC OF CEREBROSPINAL FLUID

Abstract

Experiments are described which have led to the standardization of a procedure for the purification of the γ_cglobulin 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_cglobulin in the CSF applied to the column. From an average CSF pool the first fraction, F₁, eluted by 0.002 M TRIS–HCl will usually contain only the γ_cglobulin. The second fraction F₂, eluted by 0.02 M TRIS-HCl, will contain a mixture of the γ_c, γ_s, and β_cglobulins. The third fraction F₃, eluted by 0.08 M TRIS–HCl, will contain chiefly γ_sglobulin as well as traces of the γ_cand β_cglobulins. Most of the γ_cglobulin from pooled CSF may then be obtained in purified form by rechromatographing the F₂'s from several runs. Almost all of the γ_cglobulin applied will be found in F₁. The γ_sand β_cglobulins will be eluted in the other two fractions.