Quantification of prothrombin in human plasma amplified by autocatalytic reaction
2012 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 84, no 5, 2380-2387 p.Article in journal (Refereed) Published
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.
IdentifiersURN: urn:nbn:se:umu:diva-73232DOI: 10.1021/ac203138yPubMedID: 22324776OAI: oai:DiVA.org:umu-73232DiVA: diva2:630460