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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MDPI AG
2022-01-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/12/2/313 |
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| author | Yukihiro Nishikawa Shiyo Nishida Keiko Kuroda Hirokazu Kashiwagi Yoshiaki Tomiyama Masataka Kuwana |
| author_facet | Yukihiro Nishikawa Shiyo Nishida Keiko Kuroda Hirokazu Kashiwagi Yoshiaki Tomiyama Masataka Kuwana |
| author_sort | Yukihiro Nishikawa |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T22:11:58Z |
| format | Article |
| id | doaj.art-0980dbe6d4d841eb92340f4ff34b44ae |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-09T22:11:58Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-0980dbe6d4d841eb92340f4ff34b44ae2023-11-23T19:29:56ZengMDPI AGDiagnostics2075-44182022-01-0112231310.3390/diagnostics12020313Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human PlasmaYukihiro Nishikawa0Shiyo Nishida1Keiko Kuroda2Hirokazu Kashiwagi3Yoshiaki Tomiyama4Masataka Kuwana5Medical & Biological Laboratories Co., Ltd., Nagoya 460-0008, JapanMedical & Biological Laboratories Co., Ltd., Nagoya 460-0008, JapanMedical & Biological Laboratories Co., Ltd., Nagoya 460-0008, JapanDepartment of Hematology and Oncology, Osaka University Graduate School of Medicine, Osaka 565-0871, JapanDepartment of Blood Transfusion, Osaka University Hospital, Osaka 565-0871, JapanDepartment of Allergy and Rheumatology, Nippon Medical School Graduate School of Medicine, Tokyo 113-8602, JapanPlasma 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.https://www.mdpi.com/2075-4418/12/2/313thrombopoietinimmunoassaychemiluminescentautomatedthrombocytopeniaimmune thrombocytopenia |
| spellingShingle | Yukihiro Nishikawa Shiyo Nishida Keiko Kuroda Hirokazu Kashiwagi Yoshiaki Tomiyama Masataka Kuwana Development of an Automated Chemiluminescent Enzyme Immunoassay for Measuring Thrombopoietin in Human Plasma Diagnostics thrombopoietin immunoassay chemiluminescent automated thrombocytopenia immune thrombocytopenia |
| 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 | thrombopoietin immunoassay chemiluminescent automated thrombocytopenia immune thrombocytopenia |
| url | https://www.mdpi.com/2075-4418/12/2/313 |
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