Our latest work on macromolecular cryoprotectants has been published in Biomacromolecules
There is increasing interest in developing new macromolecular cryoprotectants to improve cryopreservation outcomes, however, the criteria used for assessing cryopreservation success varies greatly between studies. In this work we critically analysed the impact of different macromolecular cryoprotectants on post-thaw cell viability and cell recovery, at multiple timepoints, for several cell lines. We found that cell viability was not a good predictor of cryopreservation outcome as it overlooked the number of cells lost during the cryopreservation process. In addition, we showed that it is essential to culture the cells for a period after thawing to allow apoptosis (programmed cell death) to initiate and complete. Considering these findings, we demonstrated that polyampholytes (an emerging class of macromolecular cryoprotectant) are effective cryoprotectants that improve both cell viability and cell recovery, compared to poly(ethylene glycol) which can produce in false positive results.
Read the paper here:
GibsonGroup developing a rapid Coronovirus test
For the past few months the group have been working with Iceni Diagnostics to develop an alternative tool for detecting the novel coronavirus (SARS-COV-2). We have discovered that coronavirus spike proteins (the 'outside of the virus') binds to specific glycans (sugars) and use this to 'capture' it and allow rapid detection using a lateral flow device. Lateral flow devices (e.g home pregnancy test) are easy to use and do not need any training nor infrastructure. We have have proven the principle of this method now, have made prototype devices and are looking to take it (quickly) to the next stage.
Pre-print (NOT PEER REVIEWED) describing the technology is here.
Glyconanomaterials to probe influenza Hemagglutinins is published in Biomacromolecules
Pathogens invade their hosts by several mechanisms including binding to glycans (sugars) on our cell surface. This binding is a crucial stage in their infection cycle and understanding these processes and developing new diagnostics and treatments. In this work we used glycoyslated nanoparticles to investigate how carbohydrate-binding proteins on the surface of influenza (hemagglutinins) bind sialic acids. We used our gold nanoparticle platform to enable easy incorporation of the sialic acids at the ends of polymers immobilised onto the gold particles, ensuring they were colloidally stable but still capable of present the sialic acids. Using a range of assays we optimized their structure to maximise outputs. We then interrogated a panel of hemaglutinins from different influenza strains, including zoonotic (species-crossing) strains. Crucially, when influenza 'jumps species' (e.g. avian to human), the nature of the glycan it binds also changes, which we were able to rapidly map. These results showed that our nanoparticle platform is a suitable tool for interrogating viral surface proteins and for helping to understand zoonosis.
Latest work published in Materials Horizons exploits new polymeric nanomaterials to modulate ice growth.
Polymerisation-induced self-assembly (PISA), a scalable and versatile method to obtain soft nano-objects is utilised for the first time to introduce ice recrystallisation inhibiting (IRI) polymer nanomaterials. Crucially we developed a method to ensure the particles are saline stable which is essential for IRI testing but current PISA formulations cannot tolerate any salt. Uniquely, we achieved this by tuning the core, enabling us to retain our IRI active corona (based on poly(vinyl alcohol)). The resulting particles showed remarkable activity, inhibiting all ice growth below 1 mg.mL-1. These results are significant as they show that Nature’s approach to hyperactivity, based upon aggregation/self-assembly, can be mimicked using polymer self-assembly.
Our recent work using a photochemical high-throughput discovery platform to identify new macromolecular cryoprotectants has been published in Macro Letters.
In this study we used a liquid handling system to produce 120 unique terpolymers using photopolymerisation with RAFT agents. These terpolymers were screened using a red blood cell freezing assay to identify the best and the worst polymer cryoprotectants, allowing us to explore the chemical space in a short time frame. Testing the hit polymers with a nucleated cell line demonstrated that the high throughput screen was able predict how well the polymers would perform as cryoprotective agents in a more complex assay. This new high throughput approach will allow us to identify new, potent macromolecular cryoprotectants.
Read the paper here:
New work on using X-rays for assessment of ice recrystallisation inhibition is published in Analyst
X-ray diffraction to probe the kinetics of ice-recrystallisation inhibition
We have a large program to discover and understand macromolecular cryoprotectants and new materials to control ice growth. This is typically assessed by end-point assays using averaged data from a small number of ice crystals. Here we make use of the University of Warwick's excellent X-ray scattering platform to show how we can use a 2-D detector on WAXS (wide-angle x-ray scattering) to monitor ice recrsytallisaton in a bulk sample enabling 100's of crystals to be sampled. This method enables easy access to kinetic information which is challenging to obtain from cryo-microscopy alone.
Professor Gibson Awarded €2 Million ERC Grant
Matt has been awarded a 5 years, €2 Million European Research Council (ERC) grant to start in 2020 called 'ICE PACK'. The aim of this is to develop materials for storing biologics in the cold chain, with the aim of making advanced therapies more available through better transport. This will follow from his previous ERC grants (Starting and Proof of concept) which studied polymers which could prevent ice growth. News story is here.
Our recent work into how macromolecular antifreeze agents interact with ice was recently published in ACS Macroletters.
Enhancement of Macromolecular Ice Recrystallisation Inhibition Activity by Exploiting Depletion Forces
We have shown that the ice recrystallisation inhibition of poly(vinyl alcohol) can be improved by the addition of other, inactive macromolecules, in this case poly(ethylene glycol). The additive causes a depletant effect in the liquid channels between ice grains, driving PVA out of solution and on to the the ice crystal surface.
These results give greater insight into the mechanism behind poly(vinyl alcohol)'s antifreeze activity and open up a new suite of tools we can use to make potent antifreeze/cryopreservation agents.