Purification and characterization of human immunoglobulin IgA1 and IgA2 isotypes from serum.
A method is described for the simultaneous purification of IgA1 and IgA2 from human serum. Ammonium sulphate precipitation, gel filtration and ion-exchange chromatography on DEAE-Sephacel yielded a partially purified IgA preparation which was separated quantitatively into IgA1 and IgA2 by affinity chromatography on jacalin-Sepharose. The IgA1 which bound to the jacalin was eluted with 0.8 M D-galactose. The IgA1 preparation was apparently homogeneous by SDS-PAGE but contained a trace of C1-inhibitor and a second protein detected by immunoelectrophoresis. The IgA2 which did not bind to the jacalin was purified to apparent homogeneity by chromatography on columns of Protein G-Sepharose, Fastflow-S Sepharose and Superose 6. Typical yields were 95% and 58% for IgA1 and IgA2 respectively or 253 mg and 24 mg per 100 ml serum. The IgA1 and IgA2 were characterised by their reactivity with isotype specific monoclonal antibodies and sensitivity to bacterial proteinases. The IgA2 preparation apparently contained both allotypes, IgA2m(1) and IgA2m(2).
Loomes
L M
LM
Department of Pathology, University of Dundee Medical School, Ninewells Hospital, Scotland, U.K.
Stewart
W W
WW
Mazengera
R L
RL
Senior
B W
BW
Kerr
M A
MA
D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006801:Humans; D007070:Immunoglobulin A / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000302:isolation & purification; D007132:Immunoglobulin Isotypes / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000302:isolation & purification
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M