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Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma

Plasma thrombopoietin (TPO) measurements help distinguish between different types of thrombocytopenia but are not feasible in routine clinical practice. We developed a fully automated quantitative chemiluminescent enzyme immunoassay (CLEIA) for measuring TPO (TPO-CLEIA), which is a one-step sandwich...

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Autores principales: Nishikawa, Yukihiro, Nishida, Shiyo, Kuroda, Keiko, Kashiwagi, Hirokazu, Tomiyama, Yoshiaki, Kuwana, Masataka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8871323/
https://www.ncbi.nlm.nih.gov/pubmed/35204403
http://dx.doi.org/10.3390/diagnostics12020313
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author Nishikawa, Yukihiro
Nishida, Shiyo
Kuroda, Keiko
Kashiwagi, Hirokazu
Tomiyama, Yoshiaki
Kuwana, Masataka
author_facet Nishikawa, Yukihiro
Nishida, Shiyo
Kuroda, Keiko
Kashiwagi, Hirokazu
Tomiyama, Yoshiaki
Kuwana, Masataka
author_sort Nishikawa, Yukihiro
collection PubMed
description Plasma thrombopoietin (TPO) measurements help distinguish between different types of thrombocytopenia but are not feasible in routine clinical practice. We developed a fully automated quantitative chemiluminescent enzyme immunoassay (CLEIA) for measuring TPO (TPO-CLEIA), which is a one-step sandwich-type assay. This assay utilizes a mouse monoclonal capture antibody, which has the neutralizing epitope of the interaction between TPO and the TPO receptor, and a newly generated rabbit monoclonal detector antibody. In analytical performance studies, this assay showed good linearity over the measuring range and high sensitivity. The limit of quantification (LoQ) of this assay was 3.4 pg/mL; low TPO concentration values of almost all healthy individuals exceeded the LoQ value. In clinical validation studies, TPO levels obtained from patients with aplastic anemia (AA) significantly increased, whereas those of patients with immune thrombocytopenia (ITP) were normal or slightly increased. The cutoff value for TPO-CLEIA corresponding to the previously reported values was useful for distinguishing between ITP and AA. These results suggest that TPO-CLEIA can quantify human plasma TPO levels with high accuracy and sensitivity and has the potential to facilitate routine clinical measurement of TPO in patients with various types of thrombocytopenia.
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spelling pubmed-88713232022-02-25 Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma Nishikawa, Yukihiro Nishida, Shiyo Kuroda, Keiko Kashiwagi, Hirokazu Tomiyama, Yoshiaki Kuwana, Masataka Diagnostics (Basel) Article Plasma thrombopoietin (TPO) measurements help distinguish between different types of thrombocytopenia but are not feasible in routine clinical practice. We developed a fully automated quantitative chemiluminescent enzyme immunoassay (CLEIA) for measuring TPO (TPO-CLEIA), which is a one-step sandwich-type assay. This assay utilizes a mouse monoclonal capture antibody, which has the neutralizing epitope of the interaction between TPO and the TPO receptor, and a newly generated rabbit monoclonal detector antibody. In analytical performance studies, this assay showed good linearity over the measuring range and high sensitivity. The limit of quantification (LoQ) of this assay was 3.4 pg/mL; low TPO concentration values of almost all healthy individuals exceeded the LoQ value. In clinical validation studies, TPO levels obtained from patients with aplastic anemia (AA) significantly increased, whereas those of patients with immune thrombocytopenia (ITP) were normal or slightly increased. The cutoff value for TPO-CLEIA corresponding to the previously reported values was useful for distinguishing between ITP and AA. These results suggest that TPO-CLEIA can quantify human plasma TPO levels with high accuracy and sensitivity and has the potential to facilitate routine clinical measurement of TPO in patients with various types of thrombocytopenia. MDPI 2022-01-26 /pmc/articles/PMC8871323/ /pubmed/35204403 http://dx.doi.org/10.3390/diagnostics12020313 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nishikawa, Yukihiro
Nishida, Shiyo
Kuroda, Keiko
Kashiwagi, Hirokazu
Tomiyama, Yoshiaki
Kuwana, Masataka
Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma
title Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma
title_full Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma
title_fullStr Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma
title_full_unstemmed Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma
title_short Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma
title_sort development of an automated chemiluminescent enzyme immunoassay for measuring thrombopoietin in human plasma
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8871323/
https://www.ncbi.nlm.nih.gov/pubmed/35204403
http://dx.doi.org/10.3390/diagnostics12020313
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