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Advancing the immunoaffinity platform AFFIRM to targeted measurements of proteins in serum in the pg/ml range
Lund Univ, Dept Immunotechnol, Lund, Sweden.;Alligator Biosci, Lund, Sweden..
Lund Univ, Dept Immunotechnol, Lund, Sweden.;Camurus, Lund, Sweden..
Lund Univ, Dept Immunotechnol, Lund, Sweden.;BioGaia AB, Lund, Sweden..
Lund Univ, Dept Immunotechnol, Lund, Sweden.;Lund Univ, Dept Clin Sci, Malmo, Sweden..
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2018 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 13, no 2, article id e0189116Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
PUBLIC LIBRARY SCIENCE , 2018. Vol. 13, no 2, article id e0189116
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Analytical Chemistry
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URN: urn:nbn:se:kth:diva-225308DOI: 10.1371/journal.pone.0189116ISI: 000425083400002PubMedID: 29438379Scopus ID: 2-s2.0-85042097260OAI: oai:DiVA.org:kth-225308DiVA, id: diva2:1195248
Note

QC 20180404

Available from: 2018-04-04 Created: 2018-04-04 Last updated: 2018-04-04Bibliographically approved

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Persson, Helena

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