Chromatographic enzyme immunoassay for T-2 toxin.
Both the active ester and maleimide moieties of the cross-linking reagent, N-[(gamma-maleimidobutyryl)oxy]succinimide (GMBS), were found to react with the primary amino groups on ribonuclease (RNase). This largely inactivated RNase towards a polymeric (but not monomeric) substrate. Citraconylating the RNase first, so that essentially only a single primary amino group remained to react with GMBS, overcame this problem. The subsequent maleimido-citraconyl-RNase was used to prepare a 1:1.1 M conjugate of anti-T-2 toxin Fab' and RNase (Fab'-RNase) in a 76% yield. The conjugate was used to detect as little as 0.1 microgram of T-2 toxin based on the ability of T-2 toxin to specifically displace Fab'-RNase complexed to a T-2 agarose affinity gel.
Department of Medicinal Chemistry in the College of Pharmacy and Allied Health Professions, Boston, MA.
B A
BA
Warden
A
A
Sentissi
M
M
Ehrat
D J
DJ
Cecchini
K
K
Alam
R W
RW
Giese
D002845:Chromatography / Q000379:methods; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments; D008301:Maleimides; D012260:Ribonucleases; D012717:Sesquiterpenes / Q000032:analysis*; D013605:T-2 Toxin / Q000032:analysis*
D007140:Immunoglobulin Fab Fragments; D008301:Maleimides; D012717:Sesquiterpenes; D012260:Ribonucleases; D013605:T-2 Toxin