A group of U.K. scientists have developed two new technologies for rapidly analyzing bioprocesses. The team, from University College London, has developed a “virus laser” and a currently unnamed system for close to real-time monitoring of products and impurities.

“This is very new work. We’ve presented results, but haven’t yet published a paper,” says John Hales, PhD, a research fellow from the department of biochemical engineering, who presented his work at the Bioprocessing Summit Europe in March.

John Hales, PhD, invented a virus laser for quantifying soluble substances, such as proteins or antibodies, during bioprocessing. The schematic illustrates an optical parametric oscillator (OPO) as the source of laser excitation, a photomultiplier tube (PMT) as a detector, and the laser emission was attenuated by neutral density (ND) filters. The image shows the emission from the virus laser (green light) as viewed from the axis of the resonator. [Credit: Creative Commons, Nature Communications, “Virus Lasers for Biological Detection,” August 9, 2019, Copyright 2019, The Author(s)]

According to Hales, the virus laser technology is a ligand-binding assay designed as an alternative to ELISA, a common assay for quantifying soluble substances, such as proteins and antibodies. The scientists are aiming to develop a set of biorecognition probes that can bind to target molecules, but also generate a laser field. The first probe was a dye-labelled M13 bacteriophage, hence the name virus laser.

Hales explains how the dye-labelled M13, when it flows into a laser photometer, generates a laser signal. Small changes in the probe concentration of 50% can lead to a 10,000 change in the output signal, he explains.

“So, what we’re looking to develop, and it is early stage, is a ligand-binding assay that could rapidly report on the concentrations of, say, product or impurities, ideally working at-line to the bioprocess,” continues Hale.

In 2020, Hales was awarded a £1.2 million UK Research and Innovation Future Leaders Fellowship to work on the virus laser.

The second technology, which is currently unnamed, aims to quantify proteins within mixtures in real-time without using labels, such as when proteins co-elute from chromatography columns.

According to Hales, the technology consists of a short-pulse nanosecond laser that emits at a UV wavelength of 266nm. The pulse is steered towards a capillary in which proteins are flowing, for example, out of a chromatography column.

The proteins are excited, and their fluorescence captured by an ellipsoidal reflector and a photodiode, Hales explains. The team then uses a sampling oscilloscope to digitize the signal and data processing algorithms to, for example, identify the different proteins.

“You have a signal that you can measure, which is directly linked to the protein structure,” he says. “And you can use that like an optical signature.”

The researchers are currently spinning out a company, Roxijen, from the university. They are also working with industrial partners.

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