Large-scale purification of staphylococcal enterotoxins A, B, and C2 by dye ligand affinity chromatography.
A simple method for the purification of staphylococcal enterotoxins A (SEA), B (SEB), and C2 (SEC2) from fermentor-grown cultures was developed. The toxins were purified by pseudo-affinity chromatography by using the triazine textile dye "Red A" and gave overall yields of 49% (SEA), 44% (SEB), and 53% (SEC2). The purified toxins were homogeneous when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but isoelectric focusing of the preparations revealed the microheterogeneity associated with these toxins. The SEA and SEB preparations each consisted of two isoelectric forms with pI values of 7.3 and 6.8 (SEA) and 8.9 and 8.55 (SEB); in contrast, SEC2 contained five different isoelectric forms, with pI values ranging between 7.6 and 6.85. The pattern of elution of the isoelectric forms from the column indicated a cationic-exchange process involved in the binding of toxin to Red A. Such a method forms the basis of a high-yielding, rapid means of purifying the staphylococcal enterotoxins that can easily be adapted to large-scale production.
Brehm
R D
RD
Biologics Division, Center for Applied Microbiology and Research, Salisbury, Wiltshire, England.
Tranter
H S
HS
Hambleton
P
P
Melling
J
J
D002846:Chromatography, Affinity / Q000379:methods*; D004396:Coloring Agents; D004768:Enterotoxins / Q000302:isolation & purification*; D007526:Isoelectric Point; D013211:Staphylococcus aureus / Q000032:analysis; D014227:Triazines
D004396:Coloring Agents; D004768:Enterotoxins; D014227:Triazines; C026066:enterotoxin A, Staphylococcal; C031818:enterotoxin B, staphylococcal; C034959:enterotoxin C, staphylococcal; C042516:reactive red 120 dye