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Selection of Transporter-Targeted Inhibitory Nanobodies by Solid-Supported-Membrane SSM-Based Electrophysiology

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Antibody Selection on Cells Targeting Membrane Proteins.

Viktor Glaser^1,2, Ümran Karsli-Ünal¹, Maike Hagedorn¹

¹Department of Gastroenterology, Hepatology, Infectious Diseases and Endocrinology, Hannover Medical School, Hannover, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|September 7, 2023

Summary

Antibody phage display on cells generates specific antibodies for cell-bound targets. This method overcomes challenges with recombinant proteins, enabling antibody development for diagnostics and therapeutics.

Keywords:

Antibody phage display Antibody selection Cell sorting Flow cytometry Membrane proteins Panning

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Area of Science:

Biotechnology
Immunology
Molecular Biology

Background:

Antibody phage display is a powerful technique for antibody generation.
Selection on immobilized proteins can be limited by protein expression challenges.
Cell-based selection offers an alternative for difficult-to-express or unavailable targets.

Purpose of the Study:

To describe a straightforward protocol for generating and screening single-chain variable fragment (scFv) binders using antibody phage display on cells.
To highlight the advantages of cell-based phage display over traditional methods.

Main Methods:

Phage display selection was performed directly on target cells.
Screening involved identifying and characterizing scFv binders.
The protocol focuses on a simplified approach for antibody generation.

Main Results:

The method allows for the generation of highly specific antibodies.
It circumvents issues associated with recombinant protein expression, such as altered folding or loss of epitopes.
This approach is particularly effective for generating antibodies against membrane proteins.

Conclusions:

Cell-based antibody phage display is an effective strategy for producing specific antibodies.
It provides a valuable tool for research, diagnostics, and therapeutic applications.
The described protocol facilitates the generation of antibodies against challenging targets.