Two methods for the introduction of amino groups into agarose-based matrices: their use in immunoaffinity chromatography.
In this study two methods of coupling antibody to Sepharose CL2B are described. They involve the introduction of amino groups either via an amino acid such as glycine or through polyethyleneimine. After introduction of amino groups into the matrices, their activation and simultaneous fixing were accomplished by treatment with glutaraldehyde. Monoclonal antibody raised against von Willebrand factor was used as a model ligand to demonstrate the stability and performance of the affinity supports. Both methods examined in this study resulted in good retention of the antibody's binding capabilities and excellent stability of the derivatized matrices. Leaching of the insolubilized protein was considerably less with the polyethyleneimine-glutaraldehyde than with cyanogen bromide.
Stubbings
D
D
Natal Institute of Immunology, Durban, Republic of South Africa.
Bubb
M O
MO
Conradie
J D
JD
D000588:Amines / Q000737:chemistry; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods*; D003488:Cyanogen Bromide; D004797:Enzyme-Linked Immunosorbent Assay; D005976:Glutaral; D005998:Glycine; D006801:Humans; D007202:Indicators and Reagents; D011094:Polyethyleneimine; D012685:Sepharose / Q000737:chemistry*; D014841:von Willebrand Factor / Q000032:analysis / Q000276:immunology
D000588:Amines; D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D014841:von Willebrand Factor; D011094:Polyethyleneimine; D012685:Sepharose; D003488:Cyanogen Bromide; D005976:Glutaral; D005998:Glycine