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Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
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A Chemical Approach for Programmable Protein Outputs Based on Engineered Cell Interactions.

Daniel A Jacome1, Justin D Northrup1,2,3, Andrew J Ruff1

  • 1Department of Radiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.

ACS Chemical Biology
|December 22, 2020
PubMed
Summary

Researchers developed a novel small-molecule system to monitor and control engineered cell-cell communication. This breakthrough enables programmable protein outputs for studying complex biological systems, particularly in immuno-oncology.

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Area of Science:

  • Synthetic Biology
  • Cellular Communication
  • Biochemistry

Background:

  • Cell-cell interactions are vital for mammalian physiology, including neurocognitive and immune functions.
  • Existing tools primarily focus on intracellular processes, with limited options for probing intercellular communication.
  • Current methods for studying cell interactions often require direct protein contact and lack tunable protein expression for diverse outputs.

Purpose of the Study:

  • To develop a novel small-molecule approach for detecting and controlling engineered cell-cell communication.
  • To create a system capable of reporting the presence of two distinct engineered cell populations with programmable protein outputs.
  • To establish a tool applicable to fields like immuno-oncology.

Main Methods:

  • Utilized a trimethoprim prodrug-enzyme pair for cell-cell reporting.
  • Employed bacterial nitroreductase enzyme catalysis, orthogonal to mammalian biology.
  • Leveraged trimethoprim diffusion from activator to receiver cells to trigger outputs.
  • Validated the system using biochemical and in vitro culture assays with optical and cytokine readouts.

Main Results:

  • Demonstrated a novel small-molecule system for engineered cell-cell communication.
  • Successfully reported the presence of two different engineered cell populations.
  • Achieved programmable protein outputs, including optical and cytokine readouts.
  • Showcased the orthogonality of the bacterial nitroreductase system within mammalian cells.

Conclusions:

  • This study presents the first small-molecule approach for detecting and controlling engineered cell-cell outputs.
  • The developed system offers a versatile platform for studying intercellular processes.
  • Anticipated future applications are particularly relevant to immuno-oncology research and development.