RT Journal Article
SR Electronic
T1 Improving oligo-conjugated antibody signal in multimodal single-cell analysis
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.15.153080
DO 10.1101/2020.06.15.153080
A1 Terkild Brink Buus
A1 Alberto Herrera
A1 Ellie Ivanova
A1 Eleni Mimitou
A1 Anthony Cheng
A1 Ramin Sedaghat Herati
A1 Thales Papagiannakopoulos
A1 Peter Smibert
A1 Niels Ødum
A1 Sergei B. Koralov
YR 2021
UL http://biorxiv.org/content/early/2021/03/13/2020.06.15.153080.abstract
AB Simultaneous measurement of surface proteins and gene expression within single cells using oligo-conjugated antibodies offers high resolution snapshots of complex cell populations. Signal from oligo-conjugated antibodies is quantified by high-throughput sequencing and is highly scalable and sensitive. In this study, we investigated the response of oligo-conjugated antibodies towards four variables: Concentration, staining volume, cell number at staining, and tissue. We find that staining with recommended antibody concentrations cause unnecessarily high background and that concentrations can be drastically reduced without loss of biological information. Reducing volume only affects antibodies targeting abundant epitopes used at low concentrations and is counteracted by reducing cell numbers. Adjusting concentrations increases signal, lowers background and reduces costs. Background signal can account for a major fraction of the total sequencing and is primarily derived from antibodies used at high concentrations. Together, this study provides new insight into the titration response and background of oligo-conjugated antibodies and offers concrete guidelines on how such panels can be improved.Impact statement Oligo-conjugated antibodies are a powerful tool but require thorough optimization to reduce background signal, increase sensitivity, and reduce sequencing costs.Competing Interest StatementPS is co-inventor of a patent related to the single cell technology utilized in this study (US provisional patent application 62/515.180).