Hyperthermophilic polymerase enabled proximity extension assay

US9902993B2 · kind B2 · utility

Assignee

• Olink Proteomics AB · SE

Inventors

• Simon Fredriksson · Stockholm, SE
• Martin Lundberg · Uppsala, SE
• Anna Eriksson · Gately Drive, SE
• Emma Rennel-Dickens · Uppsala, SE

Key dates

• Filing date: Jan 29, 2013
• Grant date: Feb 27, 2018
• Expiry date: Jun 8, 2034

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N33/542
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

The present invention relates to a proximity probe based detection assay (“proximity assay”) for an analyte in a sample, specifically a proximity probe extension assay (PEA), an in particular to an improvement in the method to reduce non-specific “background” signals, wherein the improvement comprises the use in such assays of a hyperthermophilic polymerase, said method comprising: (a) contacting said sample with at least one set of at least first and second proximity probes, which probes each comprise an analyte-binding domain and a nucleic acid domain and can simultaneously bind to the analyte; (b) allowing the nucleic acid domains of the proximity probes to interact with each other upon binding of said proximity probes to said analyte, wherein said interaction comprises the formation of a duplex; (c) extending the 3′ end of at least one nucleic acid domain of said duplex to generate an extension product, wherein the extension reaction comprises increasing the temperature of assay above room temperature and uses a polymerase enzyme which is characterised as having less than 20% of its maximal enzyme activity at 40° C. and having less than 10% of its maximal enzyme activity at 25°…