Secretion-and-capture cell-surface display for selection of target-binding proteins

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• Cell-surface display systems are crucial for protein engineering and discovery.
• Existing methods may have limitations in displaying complex or full-length proteins.
• The Secretion and Capture Technology (SECANT™) platform offers a new approach.

Purpose of the Study:

• To introduce and validate the SECANT™ platform for cell-surface protein display.
• To demonstrate the system's capability in displaying complex proteins, specifically full-length immunoglobulin G (IgG).
• To showcase the efficiency of SECANT™ in isolating specific protein-displaying cells.

Main Methods:

• Utilized in vivo biotinylation of target proteins.
• Captured biotinylated proteins on avidin-coated parent cells.
• Employed cell sorting techniques to isolate desired clones.
• Demonstrated proof-of-concept using Saccharomyces cerevisiae.

Main Results:

• Successfully displayed full-length Herceptin IgG on the surface of Saccharomyces cerevisiae cells.
• Isolated specific Herceptin IgG-displaying yeast cells from a 10,000-fold excess of cells displaying a different antibody (lysozyme-binding).
• Validated the SECANT™ platform's ability to handle complex protein display and facilitate cell selection.

Conclusions:

• The SECANT™ platform is a viable and effective system for displaying complex proteins, including full-length IgG, on living cells.
• This technology enables efficient isolation of specific antibody-displaying cells.
• SECANT™ holds potential for applications in antibody discovery, protein engineering, and synthetic biology.