{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,9]],"date-time":"2026-08-09T02:51:01Z","timestamp":1786243861848,"version":"build-2736575974"},"reference-count":50,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2022,5,5]],"date-time":"2022-05-05T00:00:00Z","timestamp":1651708800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Aberrant glycosylation of IgA1 is involved in the development of IgA nephropathy (IgAN). There are many reports of IgAN markers focusing on the glycoform of IgA1. None have been clinically applied as a routine test. In this study, we established an automated sandwich immunoassay system for detecting aberrant glycosylated IgA1, using Wisteria floribunda agglutinin (WFA) and anti-IgA1 monoclonal antibody. The diagnostic performance as an IgAN marker was evaluated. The usefulness of WFA for immunoassays was investigated by lectin microarray. A reliable standard for quantitative immunoassay measurements was designed by modifying a purified IgA1 substrate. A validation study using multiple serum specimens was performed using the established WFA-antibody sandwich automated immunoassay. Lectin microarray results showed that WFA specifically recognized N-glycans of agglutinated IgA1 in IgAN patients. The constructed IgA1 standard exhibited a wide dynamic range and high reactivity. In the validation study, serum WFA-reactive IgA1 (WFA+-IgA1) differed significantly between healthy control subjects and IgAN patients. The findings indicate that WFA is a suitable lectin that specifically targets abnormal agglutinated IgA1 in serum. We also describe an automated immunoassay system for detecting WFA+-IgA1, focusing on N-glycans.<\/jats:p>","DOI":"10.3390\/ijms23095165","type":"journal-article","created":{"date-parts":[[2022,5,5]],"date-time":"2022-05-05T13:10:26Z","timestamp":1651756226000},"page":"5165","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Development and Evaluation of a Robust Sandwich Immunoassay System Detecting Serum WFA-Reactive IgA1 for Diagnosis of IgA Nephropathy"],"prefix":"10.3390","volume":"23","author":[{"given":"Yuta","family":"Uenoyama","sequence":"first","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8791-3205","authenticated-orcid":false,"given":"Atsushi","family":"Matsuda","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazune","family":"Ohashi","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koji","family":"Ueda","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Misaki","family":"Yokoyama","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takuya","family":"Kyoutou","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kouji","family":"Kishi","sequence":"additional","affiliation":[{"name":"Bio-Reagent Material Development, Bio-Diagnostic Reagent Technology Center, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youichi","family":"Takahama","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaaki","family":"Nagai","sequence":"additional","affiliation":[{"name":"Division of Nephrology, Narita Memorial Hospital, Toyohashi 441-8029, Aichi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takaaki","family":"Ohbayashi","sequence":"additional","affiliation":[{"name":"Division of Nephrology, Narita Memorial Hospital, Toyohashi 441-8029, Aichi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Osamu","family":"Hotta","sequence":"additional","affiliation":[{"name":"Division of Internal Medicine, Hotta Osamu Clinic (HOC), Sendai 984-0013, Miyagi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideki","family":"Matsuzaki","sequence":"additional","affiliation":[{"name":"Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2036","DOI":"10.1016\/S0140-6736(16)00272-5","article-title":"Primary glomerulonephritides","volume":"387","author":"Floege","year":"2016","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1046\/j.1523-1755.2001.060003969.x","article-title":"Mesangial IgA1 in IgA nephropathy exhibits aberrant O-glycosylation: Observations in three patients","volume":"60","author":"Allen","year":"2001","journal-title":"Kidney Int."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1172\/JCI5535","article-title":"Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibody","volume":"104","author":"Tomana","year":"1999","journal-title":"J. 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