Purification of immunoglobulin M and 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
S M
SM
Lancaster University, Lancaster, United Kingdom.
Titus
J A
JA
Coico
R
R
Amin
A
A
D000818:Animals; D001202:Ascitic Fluid / Q000737:chemistry / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D006801:Humans; D006825:Hybridomas / Q000737:chemistry / Q000276:immunology; D007072:Immunoglobulin D / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D037601:Mannose-Binding Lectin / Q000737:chemistry / Q000276:immunology; D051379:Mice; D037121:Plant Lectins / Q000737:chemistry / Q000276:immunology
C045475:Griffonia simplicifolia lectins; D007072:Immunoglobulin D; D007075:Immunoglobulin M; D037601:Mannose-Binding Lectin; D037121:Plant Lectins