Hybrid Macromolecular Constructs as a Platform for Spectral Nanoprobes for Advanced Cellular Barcoding in Flow...

Area of Science:

• Bioconjugation Chemistry
• Nanotechnology
• Immunology

Background:

• Flow cytometry is crucial for immunophenotyping leukocytes.
• Current methods can face limitations in multiplexing and intersample variability.
• Developing advanced nanoprobes can enhance cellular analysis capabilities.

Purpose of the Study:

• To synthesize advanced hybrid polymer-antibody nanoprobes for fluorescent cell barcoding.
• To enable multiplexed immunophenotyping of leukocytes using flow cytometry.
• To demonstrate a novel bioconjugation technique for creating barcoded nanoprobes.

Main Methods:

• A novel bioconjugation approach was used, combining two fluorescent dyes on a copolymer precursor.
• The copolymer was conjugated to antibodies to create barcoded nanoprobes.
• These nanoprobes were applied to live cells for immunophenotyping and barcoding.

Main Results:

• Up to six distinct nanoprobes were resolved in two-dimensional cytometry analysis.
• Each nanoprobe generated a unique double fluorescence signal based on antibody-antigen binding.
• Successful live cell barcoding of clinical samples with B cell abnormalities was achieved using two detection channels.

Conclusions:

• The developed hybrid polymer-antibody nanoprobes offer a robust method for fluorescent cell barcoding.
• This technique allows for precise immunophenotyping with low intersample variability.
• The single-tube sample preparation and measurement streamline flow cytometry assays.