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Published on: February 13, 2016
Ultrafiltration membrane for electrophoretic capturing of pathogens for AFM imaging
Victor N Morozov1, Melissa Evanskey, Yian Kim Tan
1National Center for Biodefense and Infectious Diseases, George Mason University, Manassas, Virginia 20110, USA. vmorozov@gmu.edu
Abstract:
This communication describes a simple and rapid technique for electrophoretically assisted capture of phages, viruses, and other pathogens on the surface of an ultrafiltration membrane that can be considered smooth at the nanoscale. The surface was prepared by coating commercial dialysis membrane with a micrometer-thick layer of cross-linked dextran or globular proteins. To ensure strong adherence of the coating, the surface of the dialysis membrane was activated in cold plasma. It was shown that the root-mean-square roughness of the coating was well below 1 nm when the polymer solution used for coating was allowed to slowly dry through a dialysis membrane left in direct contact with mica. Relatively small viral particles (e.g., fd phages 0.7 microm long and only 3.5 nm high in the dry state) are readily visible by AFM following electrophoretic capture from suspensions containing as few as 1 x 10(6) particles/mL onto membranes prepared as described.
Insights
This study presents a new method for capturing pathogens like viruses and phages using electrophoretic techniques on ultrafiltration membranes. This rapid technique enhances pathogen detection and analysis on nanoscale surfaces.
Area of Science:
- Biotechnology
- Nanotechnology
- Analytical Chemistry
Background:
- Pathogen detection and capture are crucial for diagnostics and research.
- Existing methods may lack speed, sensitivity, or nanoscale surface control.
- Ultrafiltration membranes offer potential for pathogen capture but require surface modification.
Purpose of the Study:
- To develop a simple and rapid technique for electrophoretically assisted capture of pathogens.
- To characterize the nanoscale surface properties of modified ultrafiltration membranes.
- To demonstrate the effectiveness of the method for capturing small viral particles.
Main Methods:
- Coating commercial dialysis membranes with cross-linked dextran or globular proteins.
- Activating the membrane surface using cold plasma for enhanced coating adherence.
- Utilizing atomic force microscopy (AFM) to assess nanoscale surface roughness.
- Employing electrophoretic capture of viral particles from suspension.
Main Results:
- Achieved nanoscale smooth surfaces with root-mean-square roughness below 1 nm.
- Successfully captured small viral particles (fd phages) using the electrophoretic technique.
- Demonstrated capture from suspensions as low as 1 x 10(6) particles/mL.
- Visualized captured particles effectively using AFM.
Conclusions:
- The developed technique provides a simple, rapid, and sensitive method for pathogen capture.
- Electrophoretic assistance on nanoscale smooth membranes is effective for viral particle concentration.
- This method has potential applications in pathogen detection, diagnostics, and sample preparation.
