RT Journal Article
SR Electronic
T1 Mass spectrometry based qualification of antibodies for plasma proteomics
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 158022
DO 10.1101/158022
A1 Claudia Fredolini
A1 Sanna Byström
A1 Laura Sanchez-Rivera
A1 Marina Ioannou
A1 Davide Tamburro
A1 Rui M. Branca
A1 Janne Lehtiö
A1 Peter Nilsson
A1 Jochen M. Schwenk
YR 2017
UL http://biorxiv.org/content/early/2017/06/30/158022.abstract
AB There is a strong need for procedures that enable context and application dependent validation of antibodies. Here we describe a high-throughput approach for the detailed assessment of the selectivity of antibodies in plasma by PLasma Immunocapture Mass Spectrometry (PLIMS). The utility of PLIMS is demonstrated by determining the enrichment profiles of 157 antibodies targeting 120 proteins in EDTA plasma. Applying four classification categories (ON-target, CO-target, OFF-target and NO-target), it was found that 60% (44/60) of antibodies directed against denoted plasma proteins qualified for plasma assays. Among these, 85% (60/71) co-enriched another protein besides their respective target. As shown for several antibodies against IGFBP2, PLIMS was furthermore capable to describe known and explore novel protein complexes in plasma. In summary, PLIMS provides detailed insights into antibody selectivity in the context of plasma, thus will contribute as a valuable procedure towards to the generation of more reliable affinity-based plasma proteomics data.