The AFFInity sRM called AFFIRM is developed within CREATE Health (1). AFFIRM is unique as the first platform that combines scFv and MS for high quality reproducible protein assays. AFFIRM has the capacity for multiplexed targeted protein enrichment performed by specific recombinant scFv antibody fragments. These are coupled to magnetic beads with subsequent on-bead trypsin digestion and liquid chromatography (LC)-SRM readout in a semi-automated workflow. In this way, the sensitivity of an antibody is combined with the specificity of the SRM readout to provide assays with multiplexed measurement of target proteins with high sensitivity, reproducibility and throughput. AFFIRM provides exact protein quantification with absolute specific readout, high sensitivity, capacity for isoform-specific readout and is automated to save labor time and for precision. Most importantly, AFFIRM has capacity to be an assay format directly applicable in a clinical setting.
- Säll, A. et al. AFFIRM--a multiplexed immunoaffinity platform that combines recombinant antibody fragments and LC-SRM analysis. J Proteome Res 13, 5837–5847 (2014).