Journal article

Publication year: 2024

Pages: 1187-1201

Journal: Journal of Thrombosis and Haemostasis

Volume number: 22

Issue number: 4

ISSN: 1538-7933

eISSN: 1538-7836

Open access status: Hybrid

DOI Link: https://doi.org/10.1016/j.jtha.2023.12.030

Publisher: Elsevier

Abstract: 
Background: Current assays for the detection of (allo)antibodies to platelet antigens are often laborious and widely based on the presence of well-characterized donor platelets. Objectives: To develop an easy-to-perform, sensitive, and specific test for the detection of antibodies against platelet antigens, in particular, glycoprotein (GP) antigens, called "Recombinantly Expressed Tagged SUrface Protein" (RETSUP) assay, which does not require donor platelets. Methods: Twin-Strep-tagged GP complexes were recombinantly expressed in human embryonic kidney 293 cells after stable transfection. These cell lines were used as antigen sources in the RETSUP assay, combining cell-based and enzyme-linked immunosorbent assay-based assay procedures. The assay performance was tested with recombinant antibodies, anti-human platelet antigen (HPA) reference plasmas, and anti-HPA patient sera. Results: Human embryonic kidney 293 cell lines stably expressing either Twin-Streplabeled GPIa/IIa, GPIIb/IIIa, GPIb/IX, or GPIb/IX/V complexes or GPV as well as the distinct HPA-1, HPA-3, and HPA-5 epitopes were successfully generated. Applying the generated GP-expressing cell lines, the developed RETSUP assay proved very sensitive and specific with recombinant antibodies targeting different GPs and human plasma/ serum samples. The results of the test were not affected by the GP carrying the TwinStrep-tag or by using freshly harvested or cryopreserved cells. Conclusion: The RETSUP assay is an easy-to-perform, sensitive, and specific assay for the detection of plasma/serum antibodies to platelet GP, with performance comparable to or better than those of current state-of-the-art assays in antiplatelet antibody diagnostics. Owing to the recombinant nature of the target antigens, it can be easily adapted to detect antibodies in other antibody-mediated diseases.

Harvard Citation style: Weber, S., Arnold, J., Sachs, U. and Luppa, P. (2024) Recombinantly Expressed Tagged SUrface Protein (RETSUP) assay: a new diagnostic system for the detection of antibodies to platelets, Journal of Thrombosis and Haemostasis, 22(4), pp. 1187-1201. https://doi.org/10.1016/j.jtha.2023.12.030

APA Citation style: Weber, S., Arnold, J., Sachs, U., & Luppa, P. (2024). Recombinantly Expressed Tagged SUrface Protein (RETSUP) assay: a new diagnostic system for the detection of antibodies to platelets. Journal of Thrombosis and Haemostasis. 22(4), 1187-1201. https://doi.org/10.1016/j.jtha.2023.12.030