Glycolipids noncovalently bound to DEAE-Sephadex. Use of the complexes for purification of IgM and IgG anti-(Gal beta 1-->3 GalNAc-) antibodies.
A method for the purification by affinity of antibodies of the IgM and IgG classes against the (Gal beta 1-->3 GalNAc-) epitope has been developed. The immunoadsorbent is based on the property of DEAE-Sephadex to bind acid glycolipids bearing this epitope, by electrostatic and hydrophobic interactions in a stable form in an aqueous medium. The acid glycolipid employed was asialo-GM1 ganglioside (GA1) derivatized to produce a carboxyl function on the olefinic bond of the sphingosine moiety (GA1 acid). The DEAE-Sephadex-GA1 acid complex was used to purify the antibodies of the IgG class from serum of an immunized rabbit and of the IgM class from a human serum. The specific activities of the purified antibodies were 1,200- to 2,400-fold higher, and the antibody activities were quantitatively recovered respect to the untreated sera, in both cases. The sequential use of two immunoadsorbents: DEAE-Sephadex-ganglioside and DEAE-Sephadex-GA1 acid, allows the separation of the two classes of immunoglobulins that recognize the same sugar residues in glycolipids.
Rodriguez
P E
PE
Departamento de Quimica Biologica-CIQUIBIC, Facultad de Ciencias Quimicas-CONICET, Universidad Nacional de Cordoba, Argentina.
Barra
J L
JL
Cumar
F A
FA
D000818:Animals; D000918:Antibody Specificity; D001323:Autoantibodies / Q000302:isolation & purification*; D001665:Binding Sites; D002240:Carbohydrate Sequence; D002417:Cattle; D002855:Chromatography, Thin Layer; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005677:G(M1) Ganglioside / Q000276:immunology*; D006017:Glycolipids / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011817:Rabbits
D001323:Autoantibodies; D000939:Epitopes; D006017:Glycolipids; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D005677:G(M1) Ganglioside