Abstract
M13 bacteriophage is a well-established versatile nano-building block, which can be employed to produce novel self-assembled functional materials and devices. Sufficient production and scalability of the M13, often require a large quantity of the virus and thus, improved propagation methods characterised by high capacity and degree of purity are essential. Currently, the ‘gold-standard’ is represented by infecting Escherichia coli cultures, followed by precipitation with polyethylene glycol (PEG). However, this is considerably flawed by the accumulation of contaminant PEG inside the freshly produced stocks, potentially hampering the reactivity of the individual M13 filaments. Our study demonstrates the effectiveness of implementing an isoelectric precipitation procedure to reduce the residual PEG along with FT-IR spectroscopy as a rapid, convenient and effective analytic validation method to detect the presence of this contaminant in freshly prepared M13 stocks.
Highlights
M13 bacteriophage is a well-established versatile nano-building block, which can be employed to produce novel self-assembled functional materials and devices
The UV–Vis spectra of the purified M13 using polyethylene glycol (PEG) ( M13PEG) were compared to the ones acquired after further isoelectric precipitation ( M13IEP)
This study demonstrates ways to improve the batch propagation and purification via PEG/isoelectric precipitation of the M13 bacteriophage, further combined with an alternative and facile approach to detect the presence of the residual PEG in the newly produced samples via the FT-IR spectroscopy
Summary
M13 bacteriophage is a well-established versatile nano-building block, which can be employed to produce novel self-assembled functional materials and devices. Bulk production of highly purified M13 virions is crucial for many of the application which requires the virus to interact with other components at a molecular level such as for instance, the development of bioassays, molecular probes and novel nanostructured materials[14,16,17,18,19,20,21,22,23]. These methods do not require sophisticated equipment, the PEG precipitation is inherently disadvantageous, producing M13 stocks with a significantly high content of residual P EG36. The latter can be resuspended in PBS, deionised water (DIW) or any other buffer of choice for specific applications
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