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Thromb Haemost 1997; 78(04): 1262-1267
DOI: 10.1055/s-0038-1657725
Rapid Communication
Schattauer GmbH Stuttgart

Sensitive Measurement of Thrombopoietin by a Monoclonal Antibody Based Sandwich Enzyme-linked Immunosorbent Assay

Claudia C Folman
1   The Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam, The Netherlands
2   The Department of Hematology, Academic Medical Centre/Netherlands Cancer Institute, Amsterdam, The Netherlands
,
Albert E G K von dem Borne
1   The Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam, The Netherlands
2   The Department of Hematology, Academic Medical Centre/Netherlands Cancer Institute, Amsterdam, The Netherlands
,
Irma H J A M Rensink
1   The Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam, The Netherlands
,
Winald Gerritsen
2   The Department of Hematology, Academic Medical Centre/Netherlands Cancer Institute, Amsterdam, The Netherlands
3   The Division of Immunology, Netherlands Cancer Institute, Amsterdam, The Netherlands
,
C Ellen van der Schoot
1   The Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam, The Netherlands
2   The Department of Hematology, Academic Medical Centre/Netherlands Cancer Institute, Amsterdam, The Netherlands
,
Masja de Haas
1   The Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam, The Netherlands
,
Lucien Aarden
1   The Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 06 1996

Accepted after resubmission 26 May 1997

Publication Date:
12 July 2018 (online)

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Summary

In this report a sensitive enzyme-linked immunosorbent assay (ELISA) for the measurement of plasma thrombopoietin (Tpo) is described that is solely based on monoclonal antibodies (MoAbs).

The assay has an intra and inter-assay variance of 5-7% and 7-13%, respectively. Native and recombinant human Tpo (rhTpo) were recognized equally well, no cross reactivity with other cytokines was found and rhTpo added to plasma and serum was completely recovered. With the ELISA, Tpo concentrations in EDTA-anticoagulated plasma of all controls (n = 193) could be determined, since the limit of detection (2 ± 0.8 A.U./ml, mean ± sd) was lower than the concentration found in controls (11 ± 8 A.U./ml, mean ± sd; 2.5th-97.5th percentile: 4-32 A.U./ml). Tpo levels in serum were on average 3.4 times higher than in plasma.

We showed in vivo that Tpo is bound by platelets, as in thrombocytopenic patients (n = 5) a platelet transfusion immediately led to a drop in plasma Tpo level, whereas in patients receiving chemotherapy the induced thrombocytopenia was followed by a rise in plasma Tpo levels.

In summary, these results indicate that this ELISA is a reliable tool for Tpo measurements and is applicable for large scale studies.

 

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