A robust immunoassay for pregnancy-associated plasma protein-A2 based on analysis of circulating antigen: establishment of normal ranges in pregnancy

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A robust immunoassay for pregnancy-associated plasma protein-A2 based on analysis of circulating antigen : establishment of normal ranges in pregnancy. / Kløverpris, Søren; Gaidamauskas, Ervinas; Rasmussen, Louise Carøe Vohlander; Overgaard, Michael Toft; Kronborg, Christian; Knudsen, Ulla Breth; Christiansen, Michael; Kumar, A; Oxvig, Claus.

I: Molecular Human Reproduction, Bind 19, Nr. 11, 11.2013, s. 756-763.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Kløverpris, S, Gaidamauskas, E, Rasmussen, LCV, Overgaard, MT, Kronborg, C, Knudsen, UB, Christiansen, M, Kumar, A & Oxvig, C 2013, 'A robust immunoassay for pregnancy-associated plasma protein-A2 based on analysis of circulating antigen: establishment of normal ranges in pregnancy', Molecular Human Reproduction, bind 19, nr. 11, s. 756-763. https://doi.org/10.1093/molehr/gat047

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Kløverpris, Søren ; Gaidamauskas, Ervinas ; Rasmussen, Louise Carøe Vohlander ; Overgaard, Michael Toft ; Kronborg, Christian ; Knudsen, Ulla Breth ; Christiansen, Michael ; Kumar, A ; Oxvig, Claus. / A robust immunoassay for pregnancy-associated plasma protein-A2 based on analysis of circulating antigen : establishment of normal ranges in pregnancy. I: Molecular Human Reproduction. 2013 ; Bind 19, Nr. 11. s. 756-763.

Bibtex

@article{b21b9e24aeaf4ec3bff1d494b411b2be,
title = "A robust immunoassay for pregnancy-associated plasma protein-A2 based on analysis of circulating antigen: establishment of normal ranges in pregnancy",
abstract = "Pregnancy-associated plasma protein-A (PAPP-A) and PAPP-A2, two homologous metzincin metalloproteases, are both tightly linked to regulation within the insulin-like growth factor (IGF) system because of their specific cleavage of IGF binding proteins. Recent studies suggest that PAPP-A may be involved in clinical conditions related to unwanted cellular growth, and the circulating levels of PAPP-A is an established biomarker in prenatal screening for chromosomal abnormalities. Microarray data indicate that PAPP-A2 has potential as a biomarker for pre-eclampsia. However, well-characterized immunological methods of quantification are not available. We therefore developed monoclonal antibodies against recombinant PAPP-A2. The antibodies were epitope mapped against recombinantly expressed chimeras between PAPP-A2 and PAPP-A. Furthermore, circulating PAPP-A2 was immunoaffinity purified and characterized by sequence analysis and mass spectrometry. Unlike PAPP-A, PAPP-A2 is a noncovalent dimer in which each subunit of 1558 amino acids originates from all of the 22 predicted coding exons. A previously hypothesized variant (PAPP-E) does not exist, but low amounts of a C-terminally truncated PAPP-A2 variant was detected. A sensitive and robust ELISA for full-length PAPP-A2 was developed and used to establish normal ranges of PAPP-A2 through pregnancy. The functional sensitivity of this ELISA at 20{\%} CV was 0.08 ng/ml, and the serum concentration of PAPP-A2 was found to increase during pregnancy in agreement with placental synthesis. The existence of this assay will enable an assessment of the biomarker potential of PAPP-A2 in pre-eclampsia as well as other clinical conditions.",
keywords = "pregnancy, biomarker, PAPP-A2, pappalysin-2, ELISA",
author = "S{\o}ren Kl{\o}verpris and Ervinas Gaidamauskas and Rasmussen, {Louise Car{\o}e Vohlander} and Overgaard, {Michael Toft} and Christian Kronborg and Knudsen, {Ulla Breth} and Michael Christiansen and A Kumar and Claus Oxvig",
year = "2013",
month = "11",
doi = "10.1093/molehr/gat047",
language = "English",
volume = "19",
pages = "756--763",
journal = "Molecular Human Reproduction",
issn = "1360-9947",
publisher = "Oxford University Press",
number = "11",

}

RIS

TY - JOUR

T1 - A robust immunoassay for pregnancy-associated plasma protein-A2 based on analysis of circulating antigen

T2 - establishment of normal ranges in pregnancy

AU - Kløverpris, Søren

AU - Gaidamauskas, Ervinas

AU - Rasmussen, Louise Carøe Vohlander

AU - Overgaard, Michael Toft

AU - Kronborg, Christian

AU - Knudsen, Ulla Breth

AU - Christiansen, Michael

AU - Kumar, A

AU - Oxvig, Claus

PY - 2013/11

Y1 - 2013/11

N2 - Pregnancy-associated plasma protein-A (PAPP-A) and PAPP-A2, two homologous metzincin metalloproteases, are both tightly linked to regulation within the insulin-like growth factor (IGF) system because of their specific cleavage of IGF binding proteins. Recent studies suggest that PAPP-A may be involved in clinical conditions related to unwanted cellular growth, and the circulating levels of PAPP-A is an established biomarker in prenatal screening for chromosomal abnormalities. Microarray data indicate that PAPP-A2 has potential as a biomarker for pre-eclampsia. However, well-characterized immunological methods of quantification are not available. We therefore developed monoclonal antibodies against recombinant PAPP-A2. The antibodies were epitope mapped against recombinantly expressed chimeras between PAPP-A2 and PAPP-A. Furthermore, circulating PAPP-A2 was immunoaffinity purified and characterized by sequence analysis and mass spectrometry. Unlike PAPP-A, PAPP-A2 is a noncovalent dimer in which each subunit of 1558 amino acids originates from all of the 22 predicted coding exons. A previously hypothesized variant (PAPP-E) does not exist, but low amounts of a C-terminally truncated PAPP-A2 variant was detected. A sensitive and robust ELISA for full-length PAPP-A2 was developed and used to establish normal ranges of PAPP-A2 through pregnancy. The functional sensitivity of this ELISA at 20% CV was 0.08 ng/ml, and the serum concentration of PAPP-A2 was found to increase during pregnancy in agreement with placental synthesis. The existence of this assay will enable an assessment of the biomarker potential of PAPP-A2 in pre-eclampsia as well as other clinical conditions.

AB - Pregnancy-associated plasma protein-A (PAPP-A) and PAPP-A2, two homologous metzincin metalloproteases, are both tightly linked to regulation within the insulin-like growth factor (IGF) system because of their specific cleavage of IGF binding proteins. Recent studies suggest that PAPP-A may be involved in clinical conditions related to unwanted cellular growth, and the circulating levels of PAPP-A is an established biomarker in prenatal screening for chromosomal abnormalities. Microarray data indicate that PAPP-A2 has potential as a biomarker for pre-eclampsia. However, well-characterized immunological methods of quantification are not available. We therefore developed monoclonal antibodies against recombinant PAPP-A2. The antibodies were epitope mapped against recombinantly expressed chimeras between PAPP-A2 and PAPP-A. Furthermore, circulating PAPP-A2 was immunoaffinity purified and characterized by sequence analysis and mass spectrometry. Unlike PAPP-A, PAPP-A2 is a noncovalent dimer in which each subunit of 1558 amino acids originates from all of the 22 predicted coding exons. A previously hypothesized variant (PAPP-E) does not exist, but low amounts of a C-terminally truncated PAPP-A2 variant was detected. A sensitive and robust ELISA for full-length PAPP-A2 was developed and used to establish normal ranges of PAPP-A2 through pregnancy. The functional sensitivity of this ELISA at 20% CV was 0.08 ng/ml, and the serum concentration of PAPP-A2 was found to increase during pregnancy in agreement with placental synthesis. The existence of this assay will enable an assessment of the biomarker potential of PAPP-A2 in pre-eclampsia as well as other clinical conditions.

KW - pregnancy

KW - biomarker

KW - PAPP-A2

KW - pappalysin-2

KW - ELISA

U2 - 10.1093/molehr/gat047

DO - 10.1093/molehr/gat047

M3 - Journal article

C2 - 23804707

VL - 19

SP - 756

EP - 763

JO - Molecular Human Reproduction

JF - Molecular Human Reproduction

SN - 1360-9947

IS - 11

ER -