Single-Cell Antibody Sequencing in Atherosclerosis Research

Area of Science:

• Immunology
• Genomics
• Biotechnology

Background:

• Single cell RNA sequencing (scRNA-seq) provides transcriptomic data but has limitations in assessing cell surface protein expression, especially in immune cells.
• The correlation between mRNA levels and cell surface protein abundance is often weak, necessitating complementary methods.

Purpose of the Study:

• To describe the methodology for generating cell surface protein expression data alongside transcriptomic data using scAb-Seq.
• To highlight the utility of scAb-Seq for comprehensive cellular profiling.

Main Methods:

• Utilizes oligonucleotide-tagged monoclonal antibodies (scAb-Seq) conjugated to DNA sequences for detection.
• Employs a tripartite DNA sequence containing amplification primers, antibody barcodes, and poly(dA) sequences.
• Simultaneously detects antibody-specific DNA sequences and cDNAs in a single reaction for each cell.

Main Results:

• Enables the capture and sequencing of surface protein expression data for individual cells.
• Integrates surface protein profiles with transcriptomic data from the same cells.
• Facilitates the generation of antibody sequencing data from tissue or blood cells.

Conclusions:

• scAb-Seq significantly enhances single-cell analysis by providing parallel surface protein and transcriptomic information.
• This technique is particularly valuable for immune cell characterization where surface markers are critical.
• The availability of large antibody panels further expands the applicability of scAb-Seq.