Potential of the sulfobetaine detergent Zwittergent 3-12 as a desorbing agent in biospecific and bioselective affinity chromatography.
The isolation in our laboratories of several antigens of interest from sporulated oocysts of Eimeria species by bioselective adsorption on matrices containing immobilized antigen-specific immunoglobulins IgG was initially unsuccessful. The preparations serving as source materials for these antigens contained low levels of the zwitterionic sulfobetaine detergent, Zwittergent 3-12. Since usually immunoaffinity processes are carried out in the presence of various detergents, we were surprised, subsequently, to find this detergent to be the cause of the problem in that it prevented antigen-antibody binding. These findings led us to study the potential role of Zwittergent 3-12 as an eluting agent from matrices holding bioselectively adsorbed materials. The results of seven case studies are presented in this paper and include experiments with beta-D-galactosidase adsorbed biospecifically and bioselectively on matrices via either specific antibody or inhibitor analogue. In all cases, Zwittergent 3-12 proved to be an effective desorbing agent.
Nollstadt
K H
KH
Department of Biochemical Parasitology, Merck Sharp & Dohme Research Laboratories, Rahway, NJ 07065-0900.
Karkhanis
Y D
YD
Gnozzio
M J
MJ
Crane
M S
MS
Gurnett
A M
AM
Schmatz
D M
DM
Turner
M J
MJ
D000818:Animals; D000953:Antigens, Protozoan / Q000302:isolation & purification; D001622:Betaine / Q000031:analogs & derivatives*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004539:Eimeria / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D013501:Surface-Active Agents
D000953:Antigens, Protozoan; D013501:Surface-Active Agents; D001622:Betaine