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Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay

Anti-plasminogen antibodies (α-PLG) were previously detected in a subpopulation of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) patients, showing a relation to renal lesions and outcome. Several studies showed different proportions of α-PLG positive AAV patients, possibly...

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Autores principales: Göçeroğlu, Arda, Grenmyr, Elsa, Berden, Annelies E., Hagen, E. Christiaan, Bunch, Donna, Sommarin, Yngve, Bruijn, Jan A., Bajema, Ingeborg M., Wieslander, Jörgen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6231631/
https://www.ncbi.nlm.nih.gov/pubmed/30419041
http://dx.doi.org/10.1371/journal.pone.0207064
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author Göçeroğlu, Arda
Grenmyr, Elsa
Berden, Annelies E.
Hagen, E. Christiaan
Bunch, Donna
Sommarin, Yngve
Bruijn, Jan A.
Bajema, Ingeborg M.
Wieslander, Jörgen
author_facet Göçeroğlu, Arda
Grenmyr, Elsa
Berden, Annelies E.
Hagen, E. Christiaan
Bunch, Donna
Sommarin, Yngve
Bruijn, Jan A.
Bajema, Ingeborg M.
Wieslander, Jörgen
author_sort Göçeroğlu, Arda
collection PubMed
description Anti-plasminogen antibodies (α-PLG) were previously detected in a subpopulation of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) patients, showing a relation to renal lesions and outcome. Several studies showed different proportions of α-PLG positive AAV patients, possibly due to differences in the assays used. We here present a new, optimized α-PLG Enzyme-Linked Immuno Sorbent Assay (ELISA) and validate the presence of α-PLG in AAV. Different ELISA set-ups were tested regarding plasminogen (PLG) antigen, concentrations, coating buffers, blocking agents, and environmental conditions. Purified lysine-PLG (lys-PLG) showed better differentiation between positive samples and negative samples than glutamic acid-PLG (glu-PLG). Therefore, lys-PLG was used as coating antigen. With the optimized α-PLG ELISA we found α-PLG in 14.3% of the myeloperoxidase (MPO)-ANCA patients, whereas all our proteinase-3 (PR3)-ANCA patients tested in our new assay were negative. Concluding, in this study we have combined important technical findings and methods from previous studies to optimize the α-PLG assay, which can be used for future research purposes and will aid in uniform reporting of α-PLG status of patients.
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spelling pubmed-62316312018-11-19 Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay Göçeroğlu, Arda Grenmyr, Elsa Berden, Annelies E. Hagen, E. Christiaan Bunch, Donna Sommarin, Yngve Bruijn, Jan A. Bajema, Ingeborg M. Wieslander, Jörgen PLoS One Research Article Anti-plasminogen antibodies (α-PLG) were previously detected in a subpopulation of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) patients, showing a relation to renal lesions and outcome. Several studies showed different proportions of α-PLG positive AAV patients, possibly due to differences in the assays used. We here present a new, optimized α-PLG Enzyme-Linked Immuno Sorbent Assay (ELISA) and validate the presence of α-PLG in AAV. Different ELISA set-ups were tested regarding plasminogen (PLG) antigen, concentrations, coating buffers, blocking agents, and environmental conditions. Purified lysine-PLG (lys-PLG) showed better differentiation between positive samples and negative samples than glutamic acid-PLG (glu-PLG). Therefore, lys-PLG was used as coating antigen. With the optimized α-PLG ELISA we found α-PLG in 14.3% of the myeloperoxidase (MPO)-ANCA patients, whereas all our proteinase-3 (PR3)-ANCA patients tested in our new assay were negative. Concluding, in this study we have combined important technical findings and methods from previous studies to optimize the α-PLG assay, which can be used for future research purposes and will aid in uniform reporting of α-PLG status of patients. Public Library of Science 2018-11-12 /pmc/articles/PMC6231631/ /pubmed/30419041 http://dx.doi.org/10.1371/journal.pone.0207064 Text en © 2018 Göçeroğlu et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Göçeroğlu, Arda
Grenmyr, Elsa
Berden, Annelies E.
Hagen, E. Christiaan
Bunch, Donna
Sommarin, Yngve
Bruijn, Jan A.
Bajema, Ingeborg M.
Wieslander, Jörgen
Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay
title Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay
title_full Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay
title_fullStr Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay
title_full_unstemmed Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay
title_short Anti-plasminogen antibodies in ANCA-associated vasculitis: An optimized anti-plasminogen assay
title_sort anti-plasminogen antibodies in anca-associated vasculitis: an optimized anti-plasminogen assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6231631/
https://www.ncbi.nlm.nih.gov/pubmed/30419041
http://dx.doi.org/10.1371/journal.pone.0207064
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