By Cally Xiao and Illustration by Natalie Hadland

Unboiling egg whites is only the beginning

More than two years have passed since researchers pioneered a technique to unboil egg white proteins, resulting in a media frenzy that intrigued the public. This technique is now being applied to explore other avenues of protein research, and has the potential to be a game-changer in the pharmaceutical industry.

The overarching goal of unboiling proteins

Led by Professor Gregory Weiss at the University of California–Irvine and in collaboration with researchers at the University of Western Australia and Flinders University in Australia, the team is interested in cheaper and faster ways of producing proteins, which can be used, for example, in the pharmaceutical industry to produce antibodies for cancer treatment. “If we can accelerate protein production, then hopefully we can lower the costs of anti-cancer drugs that use proteins. That’s our dream,” explained Professor Weiss in an interview with CamBioScience.

Protein denaturing and misfolding often render the proteins inactive. Using a vortex fluid device (VFD) to generate shear stress at high rotational speeds, the team was able to untangle and refold boiled egg white proteins as a proof of principle. Not only were the proteins subjected to rotational forces, they also experienced pressure and de-pressure from tiny Faraday waves innately generated from the VFD. After VFD processing, the activity of the main protein in egg whites, lysozyme, was restored.

This technique can be used to refold and recover the activity of many other proteins, but individual parameters such as buffer salt content, rotational speed and temperature have to be adjusted for each protein. “Optimizing for each protein is not as easy as we’d like,” said Professor Weiss. The lab is currently working with industry partners and academic collaborators to optimize VFD processing for certain proteins, and they also encourage research teams around the world to try using the VFD for their own proteins of interest.

Potential applications of the VFD

“Almost all of our experiments involve making proteins and building these molecules that often don’t cooperate,” Professor Weiss commented. One of the applications of the team’s protein research is developing biosensors for early cancer detection using a patient’s blood or urine sample. The biosensors need to be able to recognize cancer biomarkers, which are also proteins. Using the VFD to process cancer biomarkers generated in the lab ensures that these proteins are folded correctly to produce the biosensors.

Professor Weiss’ team has recently shown that the VFD can also be used to accelerate enzymatic reactions. The same principle of protein folding in the VFD results in conformational changes of the enzyme, which helps the enzyme stay in its most active form and thus speeding up its reaction. “I’m really excited about using the VFD to accelerate slow enzymes that do things like synthesise complicated antibiotics,” said Professor Weiss.

Left: Professor Weiss of UC Irvine shows off the vortex fluid device the lab uses to untangle and refold proteins. A nuclear magnetic resonance glass tube would be inserted to be spun at high rotational speeds and at a 45° angle. Photo credit: Shay Meinecke for CamBioScience.

Right: Professor Weiss of UC Irvine sits with the Ig Nobel Prize awarded to his team for “partially unboiling an egg.” The Ig Nobel Prize recognises research that “first makes people laugh, and then makes people think.” Professor Weiss has been a fan of the Ig Nobel awards since he was a graduate student. Photo credit: Shay Meinecke for CamBioScience.

The importance of scientific outreach

At first, the lab established the VFD protocol for untangling and refolding several proteins, but peer reviewers were sceptical of this technique due to those proteins being “special case proteins.” Professor Weiss needed to establish this protocol for a protein everyone agrees that is difficult to work with, and decided to use boiled egg white proteins. He believes that egg white proteins are universally accepted as simple in terms of protein diversity but difficult in terms of untangling and recovering activity after boiling. “Also, breakfast was on my mind,” he added.

It is crucial for a discovery with implications in many fields of research to reach a wide audience. Professor Weiss came up with the term “unboil” for the press release to explain the technique in terms that can capture the public’s attention and get them interested in science. Since then, the modern definition of unboil has been updated. Following the initial publication, he has also started an “Ask Me Anything” thread on Reddit. The thread reached nearly 70,000 people during the few hours that Professor Weiss was actively answering questions. “As scientists, we’re rarely given opportunities to talk to so many people at once, so this was exciting,” he said. In response to all the media attention, he said that he’s “really lucky to have an amazing team of students who pulled together.”

In five to ten years, Professor Weiss hopes to see VFDs in many labs around the world as a standard lab apparatus. To this end, his group is working on developing 3D printed models of the VFD. “We’re just scratching the surface of what’s possible with the VFD in protein research,” said Professor Weiss. This technique can improve the way many labs and industries work with proteins, and is already on its way towards reaching that goal.

 

Author: Cally Xiao. (Illustration: Natalie Hadland)

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