Goat immunoglobulin purification on phosphocellulose and DEAE Affi-Gel blue.
We describe a method for the efficient purification of immunoglobulins G (IgG) to near homogeneity from goat serum. This was achieved by performing first an AS-40 fractionation on goat serum, followed by chromatography on phosphocellulose (P11) equilibrated in citrate buffer at pH 5.7. Peak I, eluted at V0 from P11, contained all IgG and the other serum proteins, except beta-globulins and most of the alpha-2-globulins, which are eluted in a second peak with 0.24 M K-phosphate in citrate buffer at pH 6.0. Peak I, concentrated and dialyzed in 20 mM K-phosphate buffer pH 8.0, was then applied onto a DEAE Affi-Gel Blue column equilibrated in the same buffer. Two peaks were obtained from this column: peak I, eluted at V0 contained a pure IgG fraction, while the other serum proteins were in peak II. We conclude that the P11 step, performed under the conditions we report here, is very useful to retain the alpha-2 and beta-globulins, which contaminate the IgG when only the DEAE Affi-Gel Blue purification step is used.
Ninfali
P
P
Istituto di Chimica Biologica G. Fornaini, Università di Urbino, Italy.
Baronciani
L
L
Rapa
S
S
Marzioni
D
D
Mannello
F
F
D000818:Animals; D002482:Cellulose / Q000031:analogs & derivatives; D002848:Chromatography, DEAE-Cellulose / Q000379:methods*; D004396:Coloring Agents; D006041:Goats; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D014227:Triazines
D004396:Coloring Agents; D007074:Immunoglobulin G; D014227:Triazines; C012842:Cibacron Blue F 3GA; D002482:Cellulose; C008881:phosphocellulose