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Efficient Microfluidic Screening Method Using a Fluorescent Immunosensor for Recombinant Protein Secretions.

Yoshihiro Ito1,2, Ryuichi Sasaki2, Sayaka Asari2

  • 1Graduate School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Kanagawa, 226-8503, Japan.

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|April 24, 2023
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Summary
This summary is machine-generated.

Researchers developed a novel high-throughput screening method for improving microbial protein production. This technique rapidly identifies enhanced bacterial strains for biopharmaceutical manufacturing, significantly reducing development time.

Keywords:
biosensor technologydroplet microfluidicshigh throughput screeningmicroorganism engineeringprotein production

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Area of Science:

  • Biotechnology
  • Microbial Engineering
  • Protein Production

Background:

  • Microbial secretory protein expression is crucial for biopharmaceutical manufacturing.
  • Developing high-yield industrial strains is often a lengthy and complex process.

Purpose of the Study:

  • To develop a simple, versatile, high-throughput screening method for identifying bacterial strains with enhanced protein secretion.
  • To accelerate the development of industrial strains for biopharmaceutical protein production.

Main Methods:

  • Encapsulating mutagenized bacterial variants in microemulsions for culturing and protein secretion.
  • Utilizing a fluorescent immunosensor (quenchbody or Q-body) to detect secreted proteins within emulsions.
  • Employing a cell sorter to select high-protein-secreting strains based on fluorescence intensity.

Main Results:

  • Demonstrated the screening method's efficacy using Corynebacterium glutamicum.
  • Successfully screened for enhanced fibroblast growth factor 9 (FGF9) producing strains.
  • Achieved a threefold increase in FGF9 secretion compared to the parent strain.

Conclusions:

  • The developed screening method is simple, versatile, and applicable to various proteins by incorporating a detection tag.
  • This method significantly shortens the development timeline for industrial strains in biopharmaceutical production.
  • The process requires minimal reagents (Q-body) and no chemical treatments or substrates.