A simple method for the biochemical purification of Ro/SS-A antigen.
In the present study, Ro/SS-A antigen has been isolated from human spleen by a two-step procedure. In the first step most of the non-antigenic material was removed by means of ammonium sulphate precipitation and ion exchange chromatography. The final purification was obtained by passing the Ro/SS-A-containing fractions twice through a Mono Q ion exchange fast protein liquid chromatography (FPLC) column. The purified antigen showed identical immunoreactivity with crude material on CIE and was composed of two polypeptides with a molecular weight of approximately 60,000 and 55,000 respectively on SDS-PAGE, both reacting on Western blotting with a panel of anti-Ro/SS-A antisera. This system permits milligrams of highly purified antigen to be obtained from grams of human spleen.
Cattedra di Immunologia Clinica, Istituto di Patologia Medica 1, Pisa, Italy.
D H
DH
Wu
A
A
Tavoni
C
C
Garzelli
R
R
Neri
C
C
Vitali
S
S
Bombardieri
D001324:Autoantigens / Q000302:isolation & purification*; D003378:Counterimmunoelectrophoresis; D006801:Humans; D008970:Molecular Weight; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D000099207:SS-A Antigen
D001324:Autoantigens; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C116006:RO60 protein, human