Development and Evaluation of a Robust Sandwich Immunoassay System Detecting Serum WFA-Reactive IgA1 for Diagnosis of IgA Nephropathy

International Journal of Molecular Sciences

Kouji Kishi¹  Osamu Hotta²  Takaaki Ohbayashi³  Masaaki Nagai³  Yuta Uenoyama⁴  Misaki Yokoyama⁴  Hideki Matsuzaki⁴  Koji Ueda⁴  Atsushi Matsuda⁴  Takuya Kyoutou⁴  Kazune Ohashi⁴  Youichi Takahama⁴ 

[1] Bio-Reagent Material Development, Bio-Diagnostic Reagent Technology Center, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan;Division of Internal Medicine, Hotta Osamu Clinic (HOC), Sendai 984-0013, Miyagi, Japan;Division of Nephrology, Narita Memorial Hospital, Toyohashi 441-8029, Aichi, Japan;Reagent Engineering, Protein Technology Group, Sysmex Corporation, Kobe 651-2271, Hyogo, Japan;

关键词: IgA nephropathy;    immunocomplex;    glyco-diagnosis;    lectin;    lectin microarray;    automated immunoassay;   

DOI  :  10.3390/ijms23095165

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown