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Force measurements on reference material

Abstract:
In infection diseases, membrane fouling, interaction with bacteria, as well as in rapid healing of wounds for example, the way proteins interact with a surface plays an important role. On a surface, they function in a different way than in solution. On a ship hull, the larvae of the barnacle will leave tiny traces of proteins to test if the surface is attractive for long-term attachment. If we get to know more about this interaction, it will be possible to develop surface conditions that are less attractive for the barnacle. Large amounts of barnacles on a ship will have a destructive effect on flow resistance and will lead to more fuel consumption. The new measuring method makes use of a modified Atomic Force Microscope: a tiny ball glued to the cantilever of the microscope will attract protein molecules.

Tracing barnacle's footprint

Enschede, Netherlands | Posted on August 19th, 2016

Force measurements

An amount of just hundreds of protein molecules will be sufficient to determine a crucial value, called the iso-electric point (pI): this is the pH-value at which the protein has net zero electric charge. The pI value says a lot about the surroundings a protein will ‘feel comfortable’ in, and to which it preferably moves. Using the AFM microscope, of which the modified tip has collected protein molecules, it is possible to perform force measurements for different pH values. The tip will be attracted or repelled, or show no movement when the pI point is reached. For these measurements, the researchers made a special reference material consisting of several layers. Using this, the effect of a number of pH-values can be tested until the pI value is found.

Paint change

The tests have been successfully performed for a number of known proteins like fibrinogen, myoglobine and bovine albumin. And returning to the barnacle: the tiny protein footprint will contain enough molecules to determine the pI value. This quantifies the ideal surface conditions, and using this knowledge, new choices can be made for e.g. the paint that is used on a ship hull.

The research has been done within the group Materials Science and Technology of Polymers of Professor Julius Vancso, in close collaboration with colleagues of A*STAR in Singapore – Prof Vancso is a Visiting Professor there as well. His group is part of UT’s MESA+ Institute for Nanotechnology.

Full bibliographic information
‘Measuring protein isoelectric points by AFM-based force spectroscopy using trace amounts of sample’ by Shifeng Guo, Xiaojing Zhu, Dominik Janczewski, Serina Siew Chen Lee, Tao He, Serena Lay Ming Teo en Julius Vancso.

doi:10.1038/nnano.2016.118

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