Nitrotyrosine in plasma of celiac disease patients as detected by a new sandwich ELISA
| Autor: | E. A. Van Straaten, L. Koster-Kamphuis, W. A. Buurman, P. Ph. Forget, J. C. A. Ter Steege |
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| Přispěvatelé: | Algemene Heelkunde, Kindergeneeskunde, RS: NUTRIM School of Nutrition and Translational Research in Metabolism |
| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
Pathology
medicine.medical_specialty Blotting Western Context (language use) Enzyme-Linked Immunosorbent Assay Biochemistry Nitric oxide chemistry.chemical_compound Mice Western blot Antibody Specificity Physiology (medical) medicine Animals Humans Nitrates biology medicine.diagnostic_test Chemistry Nitrotyrosine Antibodies Monoclonal Serum Albumin Bovine Blood Proteins Molecular biology Immunohistochemistry Blot Celiac Disease Hemocyanins biology.protein Tyrosine Antibody Keyhole limpet hemocyanin |
| Zdroj: | Free Radical Biology and Medicine, 25(8), 953-963. Elsevier Science |
| ISSN: | 0891-5849 |
| DOI: | 10.1016/s0891-5849(98)00184-1 |
| Popis: | Department of Pediatrics, Academic Hospital Maastricht, The Netherlands. W.Buurman@AH.Unimaas.NLInflammation is characterized by increased nitric oxide production. Nitrotyrosine has recently been suggested to be useful as a marker for NO-mediated tissue damage. In context of the development of an ELISA for detection of nitrotyrosine in plasma, monoclonal anti-nitrotyrosine antibodies were developed by injecting mice with nitrated keyhole limpet hemocyanin. The specificity of the antibodies was determined by binding to nitrated BSA, lack of binding to unmodified BSA, tyrosine, 3-chlorotyrosine or phenylalanine and inhibition of binding by nitrotyrosine. The antibodies developed are useful for Western blot analysis and immunohistochemical staining. Using these antibodies a nitrotyrosine sandwich ELISA was developed with a lower detection limit of approximately 0.2 nM. The intra- and interassay variance were 2.4% and 11.9%, respectively. Using this newly developed ELISA, 1.27 +/- 1.03 microM nitrotyrosine was detected in plasma samples of celiac disease patients whereas nitrotyrosine was undetectable in control samples. Elevated nitrotyrosine levels were paralleled by an increase in plasma concentrations of NO-oxidation products (NOx), nitrite and nitrate from 15.1 +/- 6.1 microM in controls to 61.0 +/- 28.2 microM in celiac disease patients. Both nitrotyrosine and NOx levels declined when the patients were on a gluten-free diet, suggesting a relation between intestinal inflammation and plasma nitrotyrosine and NOx levels. |
| Databáze: | OpenAIRE |
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