Amplification of Signal on Cell Surfaces in Molecular Cascades

• Published in: Cells (Basel, Switzerland) Vol. 12; no. 24; p. 2858
• Main Authors: Rudchenko, Sergei, Taylor, Steven, Milosavic, Nenad, Rudchenko, Maria, Wedderhoff Tissi, Betina, Mapara, Markus Y, Stojanovic, Milan N
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01-12-2023
• Subjects: Antibodies; Antigens, CD19; CD19 antigen; CD3 antigen; CD45 antigen; CD45RA antigen; Cell Membrane; Cell surface; Cellular signal transduction; DNA strand displacement cascades; flow cytometry; Health aspects; Leukocyte Common Antigens; molecular programming; nanotechnology; Oligonucleotides; Surface markers; Viral antibodies; United States; United States > US; Iowa; nanotechnology; flow cytometry; DNA strand displacement cascades; molecular programming
• DOI: 10.3390/cells12242858
• EISSN: 2073-4409

We can formulate mixtures of oligonucleotide-antibody conjugates to act as molecular cascade-based automata that analyze pairs of cell surface markers (CD markers) on individual cells in a manner consistent with the implementation of Boolean logic-for example, by producing a fluorescent label only if two markers are present. While traditional methods to characterize cells are based on transducing signals from individual cell surface markers, these cascades can be used to combine into a single signal the presence of two or even more CDs. In our original design, oligonucleotide components irreversibly flowed from one antibody to another, driven by increased hybridizations, leading to the magnitude of the final signal on each cell being determined by the surface marker that was the least abundant. This is a significant limitation to the precise labeling of narrow subpopulations, and, in order to overcome it, we changed our design to accomplish signal amplification to a more abundant cell surface marker. We show the AMPLIFY function on two examples: (1) we amplify the fluorescent label from the CD19 marker onto a fivefold more abundant CD45, and (2) we amplify broadly distributed CD45RA to a more constant marker, CD3. We expect this new function to enable the increasingly complex Boolean analysis of cell surfaces.