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Filamentous bacteriophage stability in non-aqueous media
L Olofsson1, J Ankarloo, P O Andersson
1Bioorganic and Biophysical Chemistry Laboratory, Department of Chemistry and Biomedical Sciences, University of Kalmar, SE-391 82, Kalmar, Sweden.
Chemistry & Biology
|July 14, 2001
Summary
Filamentous bacteriophage, used in cloning and display, lose infectivity in alcohols due to coat protein solvation. This mechanism differs from other inactivation forms, suggesting phage display
Area of Science:
- Molecular Biology
- Biotechnology
Background:
- Filamentous bacteriophages are utilized as cloning and phage display vectors for studying ligand-receptor interactions.
- Exposure to biphasic interfaces causes phage contraction into non-infective I- or S-forms.
Purpose of the Study:
- To investigate the inactivation mechanism of filamentous bacteriophages by various alcohols.
- To explore the potential of phage display in non-aqueous environments.
Main Methods:
- Assessing phage infectivity at different alcohol concentrations (methanol, ethanol, 1-propanol).
- Analyzing spectral properties (UV fluorescence, circular dichroism) of f1 virions.
- Testing phagemid infectivity in acetonitrile.
Main Results:
- Phage inactivation was inversely dependent on alcohol hydrophobicity.
- Loss of infectivity correlated with spectral changes in the f1 virion.
- Phagemid pG8H6 retained infectivity in 99% acetonitrile.
Conclusions:
- Alcohols inactivate filamentous phage via coat protein solvation, disrupting the capsid.
- This inactivation mechanism is distinct from I- and S-form generation.
- Phage strains exhibit solvent resistance, enabling phage display in non-aqueous media.