کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1223921 967904 2007 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Validation of an ELISA for determination of antibodies induced in monkeys against Epi-hNE4, a recombinant protein inhibitor of human neutrophil elastase
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Validation of an ELISA for determination of antibodies induced in monkeys against Epi-hNE4, a recombinant protein inhibitor of human neutrophil elastase
چکیده انگلیسی

The engineered protein inhibitor of human neutrophil elastase, Epi-hNE4, is being developed for the treatment of cystic fibrosis. Like many recombinant proteins, Epi-hNE4 may induce antibodies in pre-clinical species and in humans. The aim of this report was to validate an ELISA to assess its immunogenicity in monkeys. We have designed and optimized a classical ELISA in which Epi-hNE4 was coated directly on microtitre plates and the antibodies were detected using a secondary antibody labelled with peroxidase. We report implementation of the recent recommendations proposed for the validation of immunogenicity assessment. The cut-off point was determined by means of statistical analysis of negative samples. Linearity, reproducibility, stability and specificity were estimated using quality control samples obtained from a pool of positive samples. The method was applied to monkeys given Epi-hNE4 by inhalation. A confirmation test and a neutralization assay were developed in order to further assess positive samples. In conclusion, we present here one of the first examples of validation in application of recent recommendations [A.R. Mire-Sluis, Y.C. Barrett, V. Devanarayan, E. Koren, H. Liu, M. Maia, T. Parish, G. Scott, G. Shankar, E. Shores, S.J. Swanson, G. Taniguchi, D. Wierda, L.A. Zuckerman, J. Immunol. Methods 289 (2004) 1–16].

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmaceutical and Biomedical Analysis - Volume 43, Issue 4, 12 March 2007, Pages 1423–1429
نویسندگان
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