Your browser has javascript turned off or blocked. This will lead to some parts of our website to not work properly or at all. Turn on javascript for best performance.

The browser you are using is not supported by this website. All versions of Internet Explorer are no longer supported, either by us or Microsoft (read more here: https://www.microsoft.com/en-us/microsoft-365/windows/end-of-ie-support).

Please use a modern browser to fully experience our website, such as the newest versions of Edge, Chrome, Firefox or Safari etc.

Default user image.

Filip Ottosson

Research student

Default user image.

Advancing the immunoaffinity platform AFFIRM to targeted measurements of proteins in serum in the pg/ml range

Author

  • Anna Säll
  • Daniel Corbee
  • Sara Vikström
  • Filip Ottosson
  • Helena Persson
  • Sofia Waldemarson

Summary, in English

There is a great need for targeted protein assays with the capacity of sensitive measurements in complex samples such as plasma or serum, not the least for clinical purposes. Proteomics keeps generating hundreds of biomarker candidates that need to be transferred towards true clinical application through targeted verification studies and towards clinically applicable analysis formats. The immunoaffinity assay AFFIRM (AFFInity sRM) combines the sensitivity of recombinant single chain antibodies (scFv) for targeted protein enrichment with a specific mass spectrometry readout through selected reaction monitoring (SRM) in an automated workflow. Here we demonstrate a 100 times improved detection capacity of the assay down to pg/ml range through the use of oriented antibody immobilization to magnetic beads. This was achieved using biotin-tagged scFv coupled to streptavidin coated magnetic beads, or utilizing the FLAG tag for coupling to anti-FLAG antibody coated magnetic beads. An improved multiplexing capacity with an 11-plex setup was also demonstrated compared to a previous 3-plex setup, which is of great importance for the analysis of panels of biomarker targets.

Department/s

  • Department of Immunotechnology
  • EXODIAB: Excellence of Diabetes Research in Sweden

Publishing year

2018-02-01

Language

English

Publication/Series

PLoS ONE

Volume

13

Issue

2

Document type

Journal article

Publisher

Public Library of Science

Topic

  • Immunology in the medical area
  • Medical Laboratory and Measurements Technologies

Status

Published

ISBN/ISSN/Other

  • ISSN: 1932-6203