Nettersheim, Felix Sebastian, Armstrong, Sujit Silas, Durant, Christopher, Blanco-Dominguez, Rafael, Roy, Payel, Orecchioni, Marco, Suryawanshi, Vasantika and Ley, Klaus (2022). Titration of 124 antibodies using CITE-Seq on human PBMCs. Sci Rep, 12 (1). BERLIN: NATURE PORTFOLIO. ISSN 2045-2322
Full text not available from this repository.Abstract
Single-cell RNA-sequencing (scRNA-Seq) is widely used to characterize immune cell populations. However, mRNA levels correlate poorly with expression of surface proteins, which are well established to define immune cell types. CITE-Seq (cellular indexing of transcriptomes and epitopes by sequencing) utilizes oligonucleotide-tagged antibodies to simultaneously analyze surface phenotypes and transcriptomes. Considering the high costs of adding surface phenotyping to scRNA-Seq, we aimed to determine which of 188 tested CITE-Seq antibodies can detect their antigens on human peripheral blood mononuclear cells (PBMCs), a commonly interrogated cell population in immunology, and find the optimal concentration for staining. The recommended concentration was optimal for 76 antibodies, whereas staining quality of 7 antibodies improved when the concentration was doubled. 33 and 8 antibodies still worked well when the concentration was reduced to 1/5 or 1/25, respectively. 64 antigens were not detected at any antibody concentration. Optimizing the antibody panel by removing antibodies not able to detect their target antigens and adjusting concentrations of the remaining antibodies will improve the analysis and may reduce costs. In conclusion, our data are a resource for building an informative and cost-effective panel of CITE-Seq antibodies and use them at their optimal concentrations in future CITE-seq experiments on human PBMCs.
| Item Type: | Journal Article | ||||||||||||||||||||||||||||||||||||
| Creators: |
|
||||||||||||||||||||||||||||||||||||
| URN: | urn:nbn:de:hbz:38-685429 | ||||||||||||||||||||||||||||||||||||
| DOI: | 10.1038/s41598-022-24371-7 | ||||||||||||||||||||||||||||||||||||
| Journal or Publication Title: | Sci Rep | ||||||||||||||||||||||||||||||||||||
| Volume: | 12 | ||||||||||||||||||||||||||||||||||||
| Number: | 1 | ||||||||||||||||||||||||||||||||||||
| Date: | 2022 | ||||||||||||||||||||||||||||||||||||
| Publisher: | NATURE PORTFOLIO | ||||||||||||||||||||||||||||||||||||
| Place of Publication: | BERLIN | ||||||||||||||||||||||||||||||||||||
| ISSN: | 2045-2322 | ||||||||||||||||||||||||||||||||||||
| Language: | English | ||||||||||||||||||||||||||||||||||||
| Faculty: | Unspecified | ||||||||||||||||||||||||||||||||||||
| Divisions: | Unspecified | ||||||||||||||||||||||||||||||||||||
| Subjects: | no entry | ||||||||||||||||||||||||||||||||||||
| Uncontrolled Keywords: |
|
||||||||||||||||||||||||||||||||||||
| URI: | http://kups.ub.uni-koeln.de/id/eprint/68542 |
Downloads
Downloads per month over past year
Altmetric
Export
Actions (login required)
 |
View Item |