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Published on: February 16, 2014
Effects of the Magnetic Orientation of M13 Bacteriophage on Phage Display Selection
Shuxu Wang1,2, Noriyuki Uchida1, Kento Ueno1,2
1RIKEN Center for Emergent Matter Science, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
Abstract:
Although phage display selection using a library of M13 bacteriophage has become a powerful tool for finding peptides that bind to target materials on demand, a remaining concern of this method is the interference by the M13 main body, which is a huge filament >103 times larger than the displayed peptide, and therefore would nonspecifically adhere to the target or sterically inhibit the binding of the displayed peptide. Meanwhile, filamentous phages are known to be orientable by an external magnetic field. If M13 filaments are magnetically oriented during the library selection, their angular arrangement relative to the target surface would be changed, being expected to control the interference by the M13 main body. This study reports that the magnetic orientation of M13 filaments vertical to the target surface significantly affects the selection. When the target surface was affinitive to the M13 main body, this orientation notably suppressed the nonspecific adhesion. Furthermore, when the target surface was less affinitive to the M13 main body and intrinsically free from the nonspecific adhesion, this orientation drastically changed the population of M13 clones obtained through library selection. The method of using no chemicals but only a physical stimulus is simple, clean, and expected to expand the scope of phage display selection.
Insights
Magnetic orientation of M13 bacteriophage filaments during selection suppresses nonspecific adhesion and alters clone populations. This simple, chemical-free physical stimulus is expected to enhance phage display applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Biophysics
Background:
- Phage display selection using M13 bacteriophage is a powerful tool for identifying binding peptides.
- The M13 main body can interfere with selection through nonspecific adhesion and steric hindrance.
- Filamentous phages are known to be orientable by external magnetic fields.
Purpose of the Study:
- To investigate the effect of magnetic orientation of M13 filaments on phage display selection.
- To determine if magnetic orientation can control M13 main body interference.
- To explore the potential of this physical stimulus to expand phage display applications.
Main Methods:
- Utilizing a library of M13 bacteriophage for selection against target materials.
- Applying an external magnetic field to orient M13 filaments vertically to the target surface during selection.
- Analyzing the impact of magnetic orientation on nonspecific adhesion and the resulting M13 clone populations.
Main Results:
- Magnetic orientation of M13 filaments significantly affected the selection process.
- Vertical orientation notably suppressed nonspecific adhesion when the target surface was affinitive to the M13 main body.
- When the target surface had low affinity for the M13 main body, magnetic orientation drastically altered the M13 clone population.
Conclusions:
- Magnetic orientation of M13 filaments is an effective physical stimulus to control phage display selection.
- This method reduces nonspecific adhesion and influences the selection outcome without chemicals.
- The technique offers a simple, clean, and potentially broadens the scope of phage display applications.
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