High performance liquid chromatography immunoaffinity purification of antibodies and antibody fragments.
A rapid antibody purification method is described which combines high performance liquid chromatography (HPLC) resolution with affinity chromatography specificity. The antigen used as immobilized ligand is bound to the HPLC column matrix by reacting the amino groups of the protein with the active epoxy groups of the latter. Once the reaction has finished, the unreacted groups of the column are saturated with an appropriate scavenger. Unrelated proteins are then washed out and the specific antibodies recovered by lowering the pH of the elution buffer.
Santucci
A
A
C.R.I.S.M.A., University of Siena, Italy.
Soldani
P
P
Lozzi
L
L
Rustici
M
M
Bracci
L
L
Petreni
S
S
Neri
P
P
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001202:Ascitic Fluid / Q000032:analysis; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D007128:Immunoglobulin Fragments / Q000302:isolation & purification*; D051379:Mice; D010446:Peptide Fragments / Q000302:isolation & purification; D010928:Placental Lactogen / Q000276:immunology; D011863:Radioimmunoassay
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007106:Immune Sera; D007128:Immunoglobulin Fragments; D010446:Peptide Fragments; D010928:Placental Lactogen