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Quantification of prothrombin in human plasma amplified by autocatalytic reaction
Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB), Histology and Cell Biology.
CIC biomaGUNE, Parque Tecnológico de San Sebastián, San Sebastián, Spain.
2012 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 84, no 5, 2380-2387 p.Article in journal (Refereed) Published
Abstract [en]

By site directed mutagenesis, we have produced recombinant mutants of human and mouse prethrombin-2 which are able to convert themselves autocatalytically into α-thrombin. We also have created a new method to amplify the signal of bioanalytical assays based on the autocatalytic activation of these mutated proenzymes. The activation of the mutants by active α-thrombin triggers an autocatalytic reaction which leads to more active thrombin resulting in the amplification of the readout signal. Addition of mutated mouse prethrombin-2 into the conventional assay for prothrombin level in human plasma, employing ecarin and the fluorogenic substrate, resulted in improvement of the detection limit by 2 orders of magnitude.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2012. Vol. 84, no 5, 2380-2387 p.
National Category
Basic Medicine
Identifiers
URN: urn:nbn:se:umu:diva-73232DOI: 10.1021/ac203138yPubMedID: 22324776OAI: oai:DiVA.org:umu-73232DiVA: diva2:630460
Available from: 2013-06-19 Created: 2013-06-19 Last updated: 2017-12-06Bibliographically approved

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