Chromatographic enzyme immunoassay for T-2 toxin
| Autor: | Beverly A. Warden, Abdellah Sentissi, Kariman Alam, Roger W. Giese, Douglas J. Cecchini, Markus Ehrat |
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| Rok vydání: | 1990 |
| Předmět: |
RNase P
Stereochemistry Immunology medicine.disease_cause Immunoenzyme Techniques Maleimides Immunoglobulin Fab Fragments chemistry.chemical_compound Ribonucleases Succinimide Affinity chromatography medicine Immunology and Allergy Ribonuclease Maleimide Chromatography biology Toxin T-2 Toxin chemistry Biochemistry biology.protein Agarose Sesquiterpenes Conjugate |
| Zdroj: | Journal of Immunological Methods. 131:77-82 |
| ISSN: | 0022-1759 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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