Identification of RIP-II toxins by affinity enrichment, enzymatic digestion and LC-MS.
Anal Chem
; 87(2): 967-74, 2015 Jan 20.
Article
em En
| MEDLINE
| ID: mdl-25496503
ABSTRACT
Type 2 ribosome-inactivating protein toxins (RIP-II toxins) were enriched and purified prior to enzymatic digestion and LC-MS analysis. The enrichment of the RIP-II family of plant proteins, such as ricin, abrin, viscumin, and volkensin was based on their affinity for galactosyl moieties. A macroporous chromatographic material was modified with a galactose-terminated substituent and packed into miniaturized columns that were used in a chromatographic system to achieve up to 1000-fold toxin enrichment. The galactose affinity of the RIP-II proteins enabled their selective enrichment from water, beverages, and extracts of powder and wipe samples. The enriched fractions were digested with trypsin and RIP-II peptides were identified based on accurate mass LC-MS data. Their identities were unambiguously confirmed by LC-MS/MS product ion scans of peptides unique to each of the toxins. The LC-MS detection limit achieved for ricin target peptides was 10 amol and the corresponding detection limit for the full method was 10 fmol/mL (0.6 ng/mL). The affinity enrichment method was applied to samples from a forensic investigation into a case involving the illegal production of ricin and abrin toxins.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Extratos Vegetais
/
Tripsina
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Cromatografia de Afinidade
/
Cromatografia Líquida
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Espectrometria de Massas em Tandem
/
Proteínas Inativadoras de Ribossomos Tipo 2
/
Galactose
Tipo de estudo:
Diagnostic_studies
Limite:
Adult
/
Humans
/
Male
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article