TY - JOUR
T1 - Specificity of two monoclonal antibodies against a synthetic glycopeptide, an analogue to the hypo-galactosylated IgA1 hinge region
AU - Hiki, Yoshiyuki
AU - Hori, Hideo
AU - Yamamoto, Kouichiro
AU - Yamamoto, Yoshihiro
AU - Yuzawa, Yukio
AU - Kitaguchi, Nobuya
AU - Takahashi, Kazuo
N1 - Funding Information:
YH received a research support grant from Sumitomo Dainippon Pharma Co., Ltd.. YY received research funds from Otsuka Pharmaceutical Co., Ltd., Kyowa Hakko Kirin Co., Ltd. and Baxter Ltd.. No other conflict of interest to declare.
Funding Information:
The authors thank Dr. T. Tomiyama for producing the monoclonal antibodies. We also thank Ms. M. Sakata, Ms. A. Kamiya and Mr. H. Itoh for their superb technical assistance and data analyses. This work was supported by grants from Fujita Health University Research Foundation and by New Energy and Industrial Technology Organization (NEDO) supported by Japanese Government. The monoclonal antibody is the subject of a patent application (Domestic patent reception No.: 51302302502).
Publisher Copyright:
© 2014, The Author(s).
PY - 2015/4/1
Y1 - 2015/4/1
N2 - Increased levels of hypo-galactosylated immunoglobulin (Ig)A1 (HG-IgA1) in IgA nephropathy (IgAN) have been detected using a Helix aspersa agglutinin lectin enzyme-linked immunosorbent assay (ELISA). In this study, we developed monoclonal antibodies to evaluate the HG-IgA1 in IgA nephropathy, aiming to gain a more consistent and reproducible assay. As an analogue to the HG-IgA1 hinge region, a 19 mer synthetic peptide with five GalNAc (sHGP) residues at positions 4, 7, 9, 11 and 15 [VPST(GalNAc)PPT(GalNAc)PS(GalNAc)PS(GalNAc)TPPT (GalNAc)PSPS-NH2] was synthesized. Two monoclonal antibodies against sHGP (35A12 and 44H8) that reacted with human IgA were developed. Also, their reactivities to serum IgA from IgAN patients (n = 49), patients with other forms of kidney diseases (OKD, n = 48), and healthy controls (HC, n = 41) were evaluated using ELISA assays. The binding levels of the two monoclonal antibodies against serum IgA were significantly higher (all comparisons, p < 0.0001, Steel–Dwass non-parametric test) in IgAN patients compared to HC and OKD patients. In each individual, there was a close correlation of IgA binding levels between 35A12 and 44H8 (R2 = 0.737). These results indicate that the monoclonal antibodies recognize similar epitopes in HG IgA1, which is found predominantly in IgAN patients. The developed antibodies are proposed as a clinically useful tool for IgAN screening.
AB - Increased levels of hypo-galactosylated immunoglobulin (Ig)A1 (HG-IgA1) in IgA nephropathy (IgAN) have been detected using a Helix aspersa agglutinin lectin enzyme-linked immunosorbent assay (ELISA). In this study, we developed monoclonal antibodies to evaluate the HG-IgA1 in IgA nephropathy, aiming to gain a more consistent and reproducible assay. As an analogue to the HG-IgA1 hinge region, a 19 mer synthetic peptide with five GalNAc (sHGP) residues at positions 4, 7, 9, 11 and 15 [VPST(GalNAc)PPT(GalNAc)PS(GalNAc)PS(GalNAc)TPPT (GalNAc)PSPS-NH2] was synthesized. Two monoclonal antibodies against sHGP (35A12 and 44H8) that reacted with human IgA were developed. Also, their reactivities to serum IgA from IgAN patients (n = 49), patients with other forms of kidney diseases (OKD, n = 48), and healthy controls (HC, n = 41) were evaluated using ELISA assays. The binding levels of the two monoclonal antibodies against serum IgA were significantly higher (all comparisons, p < 0.0001, Steel–Dwass non-parametric test) in IgAN patients compared to HC and OKD patients. In each individual, there was a close correlation of IgA binding levels between 35A12 and 44H8 (R2 = 0.737). These results indicate that the monoclonal antibodies recognize similar epitopes in HG IgA1, which is found predominantly in IgAN patients. The developed antibodies are proposed as a clinically useful tool for IgAN screening.
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U2 - 10.1007/s40620-014-0118-4
DO - 10.1007/s40620-014-0118-4
M3 - Article
C2 - 25037591
AN - SCOPUS:84925938224
VL - 28
SP - 181
EP - 186
JO - Journal of Nephrology
JF - Journal of Nephrology
SN - 1121-8428
IS - 2
ER -