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Related Concept Videos

Yeast Signaling01:28

Yeast Signaling

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Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
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Protein Engineering by Yeast Surface Display
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Yeast Display Guided Selection of pH-Dependent Binders.

Jenna N Meanor1, Albert J Keung1, Balaji M Rao1,2

  • 1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 28, 2022
PubMed
Summary

Engineered antibody recycling technology enhances therapeutic half-life by utilizing pH-dependent antigen binding. This cell sorting method selects binders with strong physiological pH binding and weak acidic pH binding for improved dosing.

Keywords:
Antibody recyclingCell sortingHalf-life extensionHistidine scanningYeast surface displaypH-dependent binding

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Area of Science:

  • Biotechnology
  • Protein Engineering
  • Immunology

Background:

  • pH-dependent antigen binding is key for antibody recycling technology.
  • This technology extends therapeutic half-life, reducing dosing frequency and amount.
  • Cell sorting and display techniques are established for selecting high-affinity binders.

Purpose of the Study:

  • To describe a novel cell sorting methodology for selecting pH-dependent binding proteins.
  • To utilize yeast surface display for identifying binders with specific pH-binding profiles.
  • To enable engineering of proteins and antibodies with desired pH-dependent characteristics.

Main Methods:

  • Yeast surface display was employed for protein selection.
  • The methodology focused on selecting proteins with strong binding at physiological pH.
  • Selection criteria included weak to no binding at acidic pH.

Main Results:

  • A cell sorting methodology for pH-dependent binder selection was successfully developed.
  • The method allows for the identification of proteins with specific binding characteristics across different pH levels.
  • This approach is adaptable for various protein engineering applications.

Conclusions:

  • The described yeast surface display methodology is effective for selecting pH-dependent binding proteins.
  • This technique can be applied to engineer existing proteins or discover new pH-dependent binders.
  • The approach offers a valuable tool for developing next-generation therapeutics with enhanced antibody recycling properties.