Compared stability of Sepharose-based immunoadsorbents prepared by various activation methods.
During the use of chromatographic supports for the purification of proteins or the selective removal of substances by immunoaffinity, leakage of the antibodies immobilized on the matrix is systematically observed. When the cleansing of blood plasma by extracorporeal circulation is concerned, it is of prime importance that the immunoadsorbents exhibit an extensive chemical stability over the whole range of experimental conditions. To study and minimize this leakage, a matrix, Sepharose CL-4B, was activated by various chemical reagents and coupled to goat anti-apolipoprotein B polyclonal antibodies. Immunoadsorbents thus prepared were compared with those obtained earlier by cyanogen bromide activation. It turns out that divinyl sulphone- and tresyl chloride-activated supports lead to similar results in terms of coupling yield and adsorption capacity, but to a significant reduction in released antibodies.
Ubrich
N
N
INSERM Unité 284, Vandoeuvre-les-Nancy, France.
Hubert
P
P
Regnault
V
V
Dellacherie
E
E
Rivat
C
C
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000918:Antibody Specificity; D001055:Apolipoproteins B / Q000097:blood / Q000302:isolation & purification; D002072:Butylene Glycols; D002846:Chromatography, Affinity; D003488:Cyanogen Bromide / Q000737:chemistry*; D006041:Goats / Q000276:immunology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D007164:Immunosorbents / Q000138:chemical synthesis / Q000737:chemistry*; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry; D013450:Sulfones
D000906:Antibodies; D001055:Apolipoproteins B; D002072:Butylene Glycols; D007164:Immunosorbents; D013450:Sulfones; C032605:2,2,2-trifluoroethanesulfonyl chloride; C009873:divinyl sulfone; C035771:Sepharose CL 4B; D012685:Sepharose; C014376:1,4-bis(2,3-epoxypropoxy)butane; D003488:Cyanogen Bromide