A rapid one step purification procedure for murine IgD based on the specific affinity of Bandeiraea (Griffonia) simplicifolia-1 for N-linked carbohydrates on IgD.
The alpha-D-galactopyranosyl binding lectin from the seeds of Bandeiraea simplicifolia (a.k.a. Griffonia simplicifolia) termed BS-I, strongly reacts with murine IgD and with no other protein in ascites including all other classes of immunoglobulins as determined by immunoprecipitation, hemagglutination inhibition and affinity binding. Based on this finding, murine IgD could be rapidly purified directly from whole ascitic fluid by passage over affinity beads of BS-I linked to Sepharose 4B and subsequent elution by a buffer containing 0.1 M D-galactose. The sugar eluted product is 95-99% pure as determined by SDS-PAGE and represents 90-95% of the total IgD in the initial ascites by ELISA assay. Both monomeric and dimeric murine IgD may be purified by this procedure. Human IgD is unreactive with this lectin. Treatment of purified IgD with endoglycosidases that remove either O- or N-linked glycosides indicates that BS-I binds to IgD only via N-linked carbohydrate chains.
Oppenheim
J D
JD
Department of Microbiology, New York University Medical Center, NY 10016.
Amin
A R
AR
Thorbecke
G J
GJ
D000818:Animals; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D002846:Chromatography, Affinity; D005779:Immunodiffusion; D007072:Immunoglobulin D / Q000302:isolation & purification*; D037102:Lectins; D051379:Mice; D008807:Mice, Inbred BALB C; D037121:Plant Lectins
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; C045475:Griffonia simplicifolia lectins; D007072:Immunoglobulin D; D037102:Lectins; D037121:Plant Lectins