کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902188 | 1545793 | 2005 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A novel capture-ELISA for detection of anti-neutrophil cytoplasmic antibodies (ANCA) based on c-myc peptide recognition in carboxy-terminally tagged recombinant neutrophil serine proteases
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کلمات کلیدی
ANCAPR3P-ANCAC-ANCAMoAbBSA - BSAMPO - DFObovine serum albumin - آلبومین سرم گاوMonoclonal antibody - آنتی بادی مونوکلونالanti-neutrophil cytoplasmic antibodies - آنتی بادیهای سیتوپلاسمی ضد نوتروفیلenzyme linked immunosorbent assay - آنزیم تست ایمونوسیورسانس مرتبط استneutrophil elastase - الاستاز نوتروفیلELISA - تست الیزاCapture ELISA - ضبط الیزاCoefficient of Variation - ضریب تغییرconfidence interval - فاصله اطمینانmyeloperoxidase - میلوپراکسیداز proteinase 3 - پروتئیناز 3Wegener's granulomatosis - گرانولوماتوز Wegener
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
Testing for antineutrophil cytoplasmic antibodies (ANCA) reacting with proteinase 3 (PR3) is part of the routine diagnostic evaluation of patients with small vessel vasculitis. For PR3-ANCA detection, capture ELISAs are reported to be superior to direct ELISAs. Standard capture ELISAs, in which PR3 is anchored by anti-PR3 monoclonal antibodies (moAB), have two potential disadvantages. First, the capturing moAB may compete for epitopes recognized by some PR3-ANCA, causing occasional false-negative results. Second, the capture of recombinant PR3 mutant molecules becomes unpredictable as modifications of specific conformational epitopes may not only affect the binding of PR3-ANCA, but also the affinity of the capturing anti-PR3 moAB. Here, we describe a new capture ELISA, and its application for PR3-ANCA detection. This new assay is based on the standardized capture of a variety of different carboxy-terminally c-myc tagged recombinant ANCA target antigens using anti-c-myc coated ELISA plates. Antigen used include c-myc tagged human rPR3 variants (mature and pro-form conformations), mouse mature rPR3 and human recombinant neutrophil elastase. This new anti-c-myc-capture ELISA for PR3-ANCA detection has an intra- and inter-assay coefficient of variation of 3.6% to 7.7%, and 15.8% to 18.4%, respectively. The analytical sensitivity and specificity for PR3-ANCA positive serum samples were 93% and 100%, respectively when rPR3 with mature conformation was used as target antigen, and 83% and 100% when the pro-enzyme conformation was employed. In conclusion, this new anti-c-myc capture ELISA compares favorably to our standard capture ELISA for PR3-ANCA detection, enables the unified capture of different ANCA target antigens through binding to a c-myc tag, and allows capture of rPR3 mutants necessary for PR3-ANCA epitope mapping studies.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 307, Issues 1â2, 20 December 2005, Pages 62-72
Journal: Journal of Immunological Methods - Volume 307, Issues 1â2, 20 December 2005, Pages 62-72
نویسندگان
Augustine S. Lee, Javier D. Finkielman, Tobias Peikert, Amber M. Hummel, Margaret A. Viss, Ulrich Specks,