AbSeq Protocol Using the Nano-Well Cartridge-Based Rhapsody Platform to Generate Protein and Transcript Expression...

Area of Science:

• Biotechnology
• Genomics
• Proteomics

Background:

• Single-cell analysis is crucial for understanding cellular heterogeneity.
• Current methods often analyze either gene or protein expression, limiting comprehensive insights.
• Integrating transcriptomic and proteomic data at the single-cell level offers a more complete biological picture.

Purpose of the Study:

• To describe a protocol for high-throughput, single-cell, multi-omic analysis.
• To enable the simultaneous measurement of over 40 proteins and 400 genes per cell.
• To facilitate sample multiplexing for tracking cell origins in pooled samples.

Main Methods:

• Utilizing oligonucleotide-labeled antibodies with single-cell RNA sequencing (scRNA-seq).
• Employing the nano-well based Rhapsody platform for high-throughput processing.
• Implementing a sample multiplexing workflow for source identification.

Main Results:

• Successful combined analysis of over 40 proteins and 400 genes per cell.
• Demonstrated scalability to over 10^4 cells.
• Validated a workflow for multiplexing samples for downstream analysis.

Conclusions:

• The described protocol enables comprehensive single-cell multi-omic profiling.
• This approach enhances the depth of understanding cellular states and functions.
• The method is applicable to various biological samples, including different tissue types and donors.