Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4.
<h4>Background</h4>Microfibrillar-associated protein 4 (MFAP4) is a systemic biomarker that is significantly elevated in samples from patients suffering from hepatic cirrhosis. The protein is generally localized to elastic fibers and other connective tissue fibers in the extracellular ma...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2013-01-01
|
Series: | PLoS ONE |
Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24324779/pdf/?tool=EBI |
id |
doaj-64199ee95e5b45a48487d4a80d6b6f94 |
---|---|
record_format |
Article |
spelling |
doaj-64199ee95e5b45a48487d4a80d6b6f942021-03-04T10:11:02ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01812e8238310.1371/journal.pone.0082383Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4.Susanne Gjørup SækmoseAnders SchlosserRené HolstSofie Lock JohanssonHelle Wulf-JohanssonIda TornøeJørgen VestboKirsten Ohm KyvikTorben BaringtonUffe HolmskovGrith Lykke Sørensen<h4>Background</h4>Microfibrillar-associated protein 4 (MFAP4) is a systemic biomarker that is significantly elevated in samples from patients suffering from hepatic cirrhosis. The protein is generally localized to elastic fibers and other connective tissue fibers in the extracellular matrix (ECM), and variation in systemic MFAP4 (sMFAP4) has the potential to reflect diverse diseases with increased ECM turnover. Here, we aimed to validate an enzyme-linked immunosorbent assay (ELISA) for the measurement of sMFAP4 with an emphasis on the robustness of the assay. Moreover, we aimed to determine confounders influencing the basal sMFAP4 variability and the genetic contribution to the basal variation.<h4>Methods</h4>The sandwich ELISA was based on two monoclonal anti-MFAP4 antibodies and was optimized and calibrated with a standard of recombinant MFAP4. The importance of pre-analytical sample handling was evaluated regarding sample tube type, time, and temperature conditions. The mean value structure and variance structure was determined in a twin cohort including 1,417 Danish twins (age 18-67 years) by mixed-effect linear regression modeling.<h4>Results</h4>The practical working range of the sandwich ELISA was estimated to be 4-75 U/ml. The maximum intra- and inter-assay variation was estimated to be 8.7% and 6.6%, respectively. Sample handling and processing appeared to influence MFAP4 measurements only marginally. The average concentration of sMFAP4 in the serum was 18.9 ± 8.4 (SD) U/ml in the twin cohort (95% CI: 18.5-19.4, median sMFAP4 17.3 U/ml). The mean structure model was demonstrated to include waist-hip ratio, age, and cigarette smoking status in interactions with gender. A relatively low heritability of h(2) = 0.24 was found after applying a model including additive genetic factors and shared and non-shared environmental factors.<h4>Conclusions</h4>The described ELISA provides robust measures of the liver fibrosis marker sMFAP4. The low heritability and the relatively limited basal variation suggest that increased sMFAP4 reflects disease-induced processes.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24324779/pdf/?tool=EBI |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Susanne Gjørup Sækmose Anders Schlosser René Holst Sofie Lock Johansson Helle Wulf-Johansson Ida Tornøe Jørgen Vestbo Kirsten Ohm Kyvik Torben Barington Uffe Holmskov Grith Lykke Sørensen |
spellingShingle |
Susanne Gjørup Sækmose Anders Schlosser René Holst Sofie Lock Johansson Helle Wulf-Johansson Ida Tornøe Jørgen Vestbo Kirsten Ohm Kyvik Torben Barington Uffe Holmskov Grith Lykke Sørensen Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. PLoS ONE |
author_facet |
Susanne Gjørup Sækmose Anders Schlosser René Holst Sofie Lock Johansson Helle Wulf-Johansson Ida Tornøe Jørgen Vestbo Kirsten Ohm Kyvik Torben Barington Uffe Holmskov Grith Lykke Sørensen |
author_sort |
Susanne Gjørup Sækmose |
title |
Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. |
title_short |
Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. |
title_full |
Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. |
title_fullStr |
Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. |
title_full_unstemmed |
Enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. |
title_sort |
enzyme-linked immunosorbent assay characterization of basal variation and heritability of systemic microfibrillar-associated protein 4. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2013-01-01 |
description |
<h4>Background</h4>Microfibrillar-associated protein 4 (MFAP4) is a systemic biomarker that is significantly elevated in samples from patients suffering from hepatic cirrhosis. The protein is generally localized to elastic fibers and other connective tissue fibers in the extracellular matrix (ECM), and variation in systemic MFAP4 (sMFAP4) has the potential to reflect diverse diseases with increased ECM turnover. Here, we aimed to validate an enzyme-linked immunosorbent assay (ELISA) for the measurement of sMFAP4 with an emphasis on the robustness of the assay. Moreover, we aimed to determine confounders influencing the basal sMFAP4 variability and the genetic contribution to the basal variation.<h4>Methods</h4>The sandwich ELISA was based on two monoclonal anti-MFAP4 antibodies and was optimized and calibrated with a standard of recombinant MFAP4. The importance of pre-analytical sample handling was evaluated regarding sample tube type, time, and temperature conditions. The mean value structure and variance structure was determined in a twin cohort including 1,417 Danish twins (age 18-67 years) by mixed-effect linear regression modeling.<h4>Results</h4>The practical working range of the sandwich ELISA was estimated to be 4-75 U/ml. The maximum intra- and inter-assay variation was estimated to be 8.7% and 6.6%, respectively. Sample handling and processing appeared to influence MFAP4 measurements only marginally. The average concentration of sMFAP4 in the serum was 18.9 ± 8.4 (SD) U/ml in the twin cohort (95% CI: 18.5-19.4, median sMFAP4 17.3 U/ml). The mean structure model was demonstrated to include waist-hip ratio, age, and cigarette smoking status in interactions with gender. A relatively low heritability of h(2) = 0.24 was found after applying a model including additive genetic factors and shared and non-shared environmental factors.<h4>Conclusions</h4>The described ELISA provides robust measures of the liver fibrosis marker sMFAP4. The low heritability and the relatively limited basal variation suggest that increased sMFAP4 reflects disease-induced processes. |
url |
https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24324779/pdf/?tool=EBI |
work_keys_str_mv |
AT susannegjørupsækmose enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT andersschlosser enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT reneholst enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT sofielockjohansson enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT hellewulfjohansson enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT idatornøe enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT jørgenvestbo enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT kirstenohmkyvik enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT torbenbarington enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT uffeholmskov enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 AT grithlykkesørensen enzymelinkedimmunosorbentassaycharacterizationofbasalvariationandheritabilityofsystemicmicrofibrillarassociatedprotein4 |
_version_ |
1714806580758708224 |