Research ExplorerPublicationsDevelopment of a standardized ELISA for the determination of...

Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy

Standard

Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy. / Dähnrich, Cornelia; Komorowski, Lars; Probst, Christian; Seitz-Polski, Barbara; Esnault, Vincent; Wetzels, Jack F; Hofstra, Julia M; Hoxha, Elion; Stahl, Rolf A; Lambeau, Gérard; Stöcker, Winfried; Schlumberger, Wolfgang.

In: CLIN CHIM ACTA, Vol. 421, 05.06.2013, p. 213-8.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Dähnrich, C, Komorowski, L, Probst, C, Seitz-Polski, B, Esnault, V, Wetzels, JF, Hofstra, JM, Hoxha, E, Stahl, RA, Lambeau, G, Stöcker, W & Schlumberger, W 2013, 'Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy', CLIN CHIM ACTA, vol. 421, pp. 213-8. https://doi.org/10.1016/j.cca.2013.03.015

APA

Dähnrich, C., Komorowski, L., Probst, C., Seitz-Polski, B., Esnault, V., Wetzels, J. F., Hofstra, J. M., Hoxha, E., Stahl, R. A., Lambeau, G., Stöcker, W., & Schlumberger, W. (2013). Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy. CLIN CHIM ACTA, 421, 213-8. https://doi.org/10.1016/j.cca.2013.03.015

Vancouver

Dähnrich C, Komorowski L, Probst C, Seitz-Polski B, Esnault V, Wetzels JF et al. Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy. CLIN CHIM ACTA. 2013 Jun 5;421:213-8. https://doi.org/10.1016/j.cca.2013.03.015

Bibtex

@article{703432d850194d0588368507fc0baf08,
title = "Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy",
abstract = "BACKGROUND: Autoantibodies against the M-type phospholipase A2 receptor (PLA2R1) are specific markers for primary membranous nephropathy (pMN) and anti-PLA2R1 serum levels may be useful to monitor disease activity. So far, a recombinant cell-based indirect immunofluorescence assay (RC-IFA) using recombinant PLA2R1 as a substrate has been widely available but lacks a finely graduated assessment of antibody concentrations.METHODS: In order to setup a standardized ELISA, the extracellular domain of human PLA2R1 was expressed in HEK293. The purified protein was used to form the solid-phase in an ELISA which was then employed to analyze sera from 200 patients with primary MN, 27 patients with secondary MN, 230 patients with other glomerular diseases, 316 patients with systemic autoimmune diseases, and from 291 healthy blood donors.RESULTS: At a set specificity of 99.9% the sensitivity of the anti-PLA2R1 IgG ELISA was found to be 96.5%. A similar sensitivity (98.5%) was obtained when binding of only subclass IgG4 was analyzed. The calibrated assay showed a good class correlation with the results of the RC-IFA, was robust and could be stored for several months without any loss of quality.CONCLUSION: The results demonstrate that the new test system is qualified for routine use and that it has an almost perfect agreement with both, the clinical characterization of the patients and the results generated with RC-IFA.",
keywords = "Adolescent, Adult, Aged, Aged, 80 and over, Autoantibodies, Biological Markers, Case-Control Studies, Enzyme-Linked Immunosorbent Assay, Female, Gene Expression, Glomerulonephritis, Membranous, HEK293 Cells, Humans, Immunoglobulin G, Male, Middle Aged, Protein Structure, Tertiary, Receptors, Phospholipase A2, Sensitivity and Specificity",
author = "Cornelia D{\"a}hnrich and Lars Komorowski and Christian Probst and Barbara Seitz-Polski and Vincent Esnault and Wetzels, {Jack F} and Hofstra, {Julia M} and Elion Hoxha and Stahl, {Rolf A} and G{\'e}rard Lambeau and Winfried St{\"o}cker and Wolfgang Schlumberger",
note = "Copyright {\textcopyright} 2013 Elsevier B.V. All rights reserved.",
year = "2013",
month = jun,
day = "5",
doi = "10.1016/j.cca.2013.03.015",
language = "English",
volume = "421",
pages = "213--8",
journal = "CLIN CHIM ACTA",
issn = "0009-8981",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Development of a standardized ELISA for the determination of autoantibodies against human M-type phospholipase A2 receptor in primary membranous nephropathy

AU - Dähnrich, Cornelia

AU - Komorowski, Lars

AU - Probst, Christian

AU - Seitz-Polski, Barbara

AU - Esnault, Vincent

AU - Wetzels, Jack F

AU - Hofstra, Julia M

AU - Hoxha, Elion

AU - Stahl, Rolf A

AU - Lambeau, Gérard

AU - Stöcker, Winfried

AU - Schlumberger, Wolfgang

N1 - Copyright © 2013 Elsevier B.V. All rights reserved.

PY - 2013/6/5

Y1 - 2013/6/5

N2 - BACKGROUND: Autoantibodies against the M-type phospholipase A2 receptor (PLA2R1) are specific markers for primary membranous nephropathy (pMN) and anti-PLA2R1 serum levels may be useful to monitor disease activity. So far, a recombinant cell-based indirect immunofluorescence assay (RC-IFA) using recombinant PLA2R1 as a substrate has been widely available but lacks a finely graduated assessment of antibody concentrations.METHODS: In order to setup a standardized ELISA, the extracellular domain of human PLA2R1 was expressed in HEK293. The purified protein was used to form the solid-phase in an ELISA which was then employed to analyze sera from 200 patients with primary MN, 27 patients with secondary MN, 230 patients with other glomerular diseases, 316 patients with systemic autoimmune diseases, and from 291 healthy blood donors.RESULTS: At a set specificity of 99.9% the sensitivity of the anti-PLA2R1 IgG ELISA was found to be 96.5%. A similar sensitivity (98.5%) was obtained when binding of only subclass IgG4 was analyzed. The calibrated assay showed a good class correlation with the results of the RC-IFA, was robust and could be stored for several months without any loss of quality.CONCLUSION: The results demonstrate that the new test system is qualified for routine use and that it has an almost perfect agreement with both, the clinical characterization of the patients and the results generated with RC-IFA.

AB - BACKGROUND: Autoantibodies against the M-type phospholipase A2 receptor (PLA2R1) are specific markers for primary membranous nephropathy (pMN) and anti-PLA2R1 serum levels may be useful to monitor disease activity. So far, a recombinant cell-based indirect immunofluorescence assay (RC-IFA) using recombinant PLA2R1 as a substrate has been widely available but lacks a finely graduated assessment of antibody concentrations.METHODS: In order to setup a standardized ELISA, the extracellular domain of human PLA2R1 was expressed in HEK293. The purified protein was used to form the solid-phase in an ELISA which was then employed to analyze sera from 200 patients with primary MN, 27 patients with secondary MN, 230 patients with other glomerular diseases, 316 patients with systemic autoimmune diseases, and from 291 healthy blood donors.RESULTS: At a set specificity of 99.9% the sensitivity of the anti-PLA2R1 IgG ELISA was found to be 96.5%. A similar sensitivity (98.5%) was obtained when binding of only subclass IgG4 was analyzed. The calibrated assay showed a good class correlation with the results of the RC-IFA, was robust and could be stored for several months without any loss of quality.CONCLUSION: The results demonstrate that the new test system is qualified for routine use and that it has an almost perfect agreement with both, the clinical characterization of the patients and the results generated with RC-IFA.

KW - Adolescent

KW - Adult

KW - Aged

KW - Aged, 80 and over

KW - Autoantibodies

KW - Biological Markers

KW - Case-Control Studies

KW - Enzyme-Linked Immunosorbent Assay

KW - Female

KW - Gene Expression

KW - Glomerulonephritis, Membranous

KW - HEK293 Cells

KW - Humans

KW - Immunoglobulin G

KW - Male

KW - Middle Aged

KW - Protein Structure, Tertiary

KW - Receptors, Phospholipase A2

KW - Sensitivity and Specificity

U2 - 10.1016/j.cca.2013.03.015

DO - 10.1016/j.cca.2013.03.015

M3 - SCORING: Journal article

C2 - 23541686

VL - 421

SP - 213

EP - 218

JO - CLIN CHIM ACTA

JF - CLIN CHIM ACTA

SN - 0009-8981

ER -