Abstract
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.
Highlights
Thrombopoietin (TPO) is the primary regulator of platelet production and maturation of megakaryocytes [1]
Many previous studies have suggested that TPO measurement is helpful in this differential diagnosis since plasma TPO levels are markedly elevated in patients with AA, while they are slightly or not elevated in patients with Immune thrombocytopenia (ITP) [13,14,15,16,17,18]
We developed a fully automated chemiluminescent enzyme immunoassay (CLEIA) based on monoclonal antibodies for measuring TPO (TPO-CLEIA)
Summary
Thrombopoietin (TPO) is the primary regulator of platelet production and maturation of megakaryocytes [1]. Previous studies have shown that blood TPO levels are inversely proportional to the number of megakaryocytes in the bone marrow and the circulating platelet count [2,3,4]. Platelet desialylation has been shown to play an important role in platelet destruction and impaired thrombopoiesis in patients with ITP [11]. Despite this progress in understanding pathophysiology, the diagnosis of ITP is still primarily based on differential diagnosis [12]. Many previous studies have suggested that TPO measurement is helpful in this differential diagnosis since plasma TPO levels are markedly elevated in patients with AA, while they are slightly or not elevated in patients with ITP [13,14,15,16,17,18]
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