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Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients
AIMS: IgA nephropathy, the most common primary glomerulonephritis worldwide, is characterized by glomerular deposition of galactose-deficient IgA1 and elevated serum levels of this IgA1 glycoform. Current ELISA methods lack sensitivity to assess galactose deficiency using small amounts of IgA1, whic...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152805/ https://www.ncbi.nlm.nih.gov/pubmed/30091383 http://dx.doi.org/10.2144/btn-2018-0042 |
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author | Reily, Colin Rizk, Dana V Julian, Bruce A Novak, Jan |
author_facet | Reily, Colin Rizk, Dana V Julian, Bruce A Novak, Jan |
author_sort | Reily, Colin |
collection | PubMed |
description | AIMS: IgA nephropathy, the most common primary glomerulonephritis worldwide, is characterized by glomerular deposition of galactose-deficient IgA1 and elevated serum levels of this IgA1 glycoform. Current ELISA methods lack sensitivity to assess galactose deficiency using small amounts of IgA1, which limits studies in primary cells due to modest IgA1 production in isolated peripheral-blood lymphocytes. METHODS: Lectin from Helix pomatia was conjugated to biotin or acridinium ester and used in ELISA to detect galactose deficiency of IgA1 using small amounts of IgA1. RESULTS: Lectin conjugated to acridinium had an approximately log-fold increased sensitivity compared with biotin-labeled lectin. CONCLUSIONS: The new method of using lectin from Helix pomatia conjugated to acridinium increased assay sensitivity, allowing future mechanistic studies with cultured primary cells. |
format | Online Article Text |
id | pubmed-6152805 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
record_format | MEDLINE/PubMed |
spelling | pubmed-61528052018-09-24 Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients Reily, Colin Rizk, Dana V Julian, Bruce A Novak, Jan Biotechniques Article AIMS: IgA nephropathy, the most common primary glomerulonephritis worldwide, is characterized by glomerular deposition of galactose-deficient IgA1 and elevated serum levels of this IgA1 glycoform. Current ELISA methods lack sensitivity to assess galactose deficiency using small amounts of IgA1, which limits studies in primary cells due to modest IgA1 production in isolated peripheral-blood lymphocytes. METHODS: Lectin from Helix pomatia was conjugated to biotin or acridinium ester and used in ELISA to detect galactose deficiency of IgA1 using small amounts of IgA1. RESULTS: Lectin conjugated to acridinium had an approximately log-fold increased sensitivity compared with biotin-labeled lectin. CONCLUSIONS: The new method of using lectin from Helix pomatia conjugated to acridinium increased assay sensitivity, allowing future mechanistic studies with cultured primary cells. 2018-08 /pmc/articles/PMC6152805/ /pubmed/30091383 http://dx.doi.org/10.2144/btn-2018-0042 Text en http://creativecommons.org/licenses/by/4.0/ This work is licensed under the Creative Commons Attribution 4.0 License. To view a copy of this license, visit http://creative-commons.org/licenses/by/4.0/ |
spellingShingle | Article Reily, Colin Rizk, Dana V Julian, Bruce A Novak, Jan Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients |
title | Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients |
title_full | Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients |
title_fullStr | Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients |
title_full_unstemmed | Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients |
title_short | Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients |
title_sort | assay for galactose-deficient iga1 enables mechanistic studies with primary cells from iga nephropathy patients |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152805/ https://www.ncbi.nlm.nih.gov/pubmed/30091383 http://dx.doi.org/10.2144/btn-2018-0042 |
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