Isolation of murine and human immunoglobulin m and murine immunoglobulin D.
This unit describes two classical protocols for the purification of IgM-dialysis of ascites fluid, tissue culture medium, or bioreactor supernatants against distilled water to precipitate pure IgM, and ammonium sulfate precipitation. Both protocols can be followed by size-exclusion chromatography to obtain a highly purified product. Recently, an affinity method for purification of IgM has been developed using mannan-binding protein, and is described here. The third approach presented is a one-step IgD purification method, designed specifically for murine derived samples, that uses Sepharose coupled to lectin derived from the seeds of Griffonia simplicifolia-1. This represents a simple, rapid, and gentle, approach to isolating this highly labile immunoglobulin from IgD-containing ascites or hybridoma sources.
Andrew
Sarah M
SM
Lancaster University, Lancaster, United Kingdom.
Titus
Julie A
JA
National Cancer Institute, Bethesda, Maryland.
Amin
Ashok
A
Carilion Clinic, Roanoke, Virginia.
Coico
Richard
R
Department of Microbiology and Immunology, Temple University School of Medicine, Philadelphia, Pennsylvania.
D000645:Ammonium Sulfate; D000818:Animals; D001202:Ascitic Fluid; D018929:Cell Culture Techniques; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D002847:Chromatography, Agarose; D002850:Chromatography, Gel; D006801:Humans; D006825:Hybridomas; D007072:Immunoglobulin D / Q000276:immunology / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000276:immunology / Q000302:isolation & purification*; D037601:Mannose-Binding Lectin; D051379:Mice; D037121:Plant Lectins
C045475:Griffonia simplicifolia lectins; D007072:Immunoglobulin D; D007075:Immunoglobulin M; D037601:Mannose-Binding Lectin; D037121:Plant Lectins; D000645:Ammonium Sulfate