Invited Speaker Details

Emerging Pharmaceutical Products Using Electric Fields

Dr. Zeeshan Ahmed, Pharmaceutical Science, University of Manchester, UK.

Zeeshan obtained both his first degree (Pharmaceutical Chemistry, with Honours ) and his PhD (EPSRC Biomedical Materials IRC) from Queen Mary, University of London. He has held several prestigious Fellowships (EPSRC, The Leverhulme Trust and The Royal Society) and has worked in several Universities (Queen Mary University of London, University College London (UCL), University of Portsmouth, De Montfort University and The University of Manchester) which includes numerous Schools of Pharmacy in the UK.

Molecular Physics of Monoclonal Antibody Formulations: Stability, Aggregation and Interfacial Adsorption

Prof. Fernando Bresme, Chemical Physics, Imperial College London, UK.

Fernando Bresme is Professor of Chemical Physics at Imperial College London, where he leads the Computational Chemical Physics Group. His research combines theory, computation and non-equilibrium thermodynamics to elucidate chemical and biological interfaces, biomolecular systems, and nanoscale transport phenomena, with applications in therapeutics and advanced materials for biomedicine.

He is a Fellow of the Royal Society of Chemistry and the Institute of Physics. His work has been recognised by an EPSRC Leadership Fellowship, the McBain Medal in Colloid and Interface Science, and election to the Royal Norwegian Academy of Sciences and Letters.

Physics, Biology and Pharmacy meet in the lungs

Prof. Pietro Cicuta, Biological Physics, Cavendish Laboratory, University of Cambridge, UK

The ciliated epithelium of the human respiratory tract is covered by the airway surface liquid (ASL), a protective fluid consisting of two layers: the periciliary layer (PCL), where motile cilia reside and generate fluid flow, and an overlying mucus layer. Beyond the mucus is a more homogeneous fluid, which is air in the human adult but a liquid during embryonic development. An effective mucus clearance is crucial to health, and restoring that clearance is the objective in pharmacological treatments. We might be able to tune better treatments if we understood better the factors that play together in enabling clearance.

Pietro Cicuta was awarded a Laurea in Physics at University of Milan in 1999, and a PhD at University of Cambridge in 2003. His research started in soft matter physics, fluids and liquid interface phenomena, then progressed to lipid membranes in postdoc and research Fellowship. He is now a Professor of Biological Physics at Cambridge University, and addresses various aspects of living systems with his research group. The group is both experiment-driven, developing bespoke tools and analysis pipelines, and computation-driven, seeking to understand biological phenomena. We employ computation and machine learning, developing new physically-grounded models to explain our results. Pietro has published over 170 peer-reviewed papers so far in his career, of which 28 are on the science of motile cilia, and 15 others are on aspects of cell biology, immunity, and epithelial infection.

Emergent properties and antibiotic tolerance of bacterial biofilms

Prof. Knut Drescher, Biozentrum, University of Basel, Switzerland.

In nature, bacteria often live in three-dimensional communities termed biofilms, in which cells are attached to each other through an extracellular matrix. These bacterial biofilms are the simplest and most ancestral forms of multicellular organization of life, and represent a useful model system for studying fundamental aspects of multicellularity. Furthermore, bacterial biofilms cause chronic infections because bacteria that are bound in biofilms are highly tolerant to antibiotic treatment. In this presentation, I will first introduce microscopy, image processing, and spatiotemporal transcriptome measurement techniques that enable us to monitor individual cells in living biofilms. By combining these techniques with antibiotic biosensors that can report on the spatiotemporal concentrations of unmodified antibiotics, I will demonstrate the dynamics of antibiotic penetration into biofilms. I will then proceed to discuss how individual cells in biofilms coordinate their activities so that the biofilm community develops emergent functions, such as protection from antibiotics or viral predators (bacteriophages). This talk will therefore shed light on the spatiotemporal development of bacterial communities, and the mechanisms underlying emergent functions of these communities.

Knut Drescher studied physics at the University of Oxford, before pursuing a PhD in biophysics at the University of Cambridge, where he studied physical aspects of the evolution of multicellularity in algal model systems. He became interested in microbiology and bacterial biofilms during his postdoc at Princeton University from 2011-2014. In 2014, Knut Drescher started his own lab at the Max Planck Institute for Terrestrial Microbiology in Germany, where he focused on combining methods from physics and molecular biology to understand the development and function of bacterial biofilms. In 2021, Knut moved his lab to the Biozentrum at the University of Basel, and continues to combine methods from physics and molecular biology to understand bacterial biofilms.

Understanding next generation diagnostic & therapeutic nanoparticles using SPARTA®

Dr. Alexandra Hopkins, Department of Physiology, Anatomy and Genetics, University of Oxford, UK.

Professor Dame Molly Stevens’ group has developed a novel platform called SPARTA® (Single Particle Automated Raman Trapping Analysis), which has now been commercialised by the spin-out company SPARTA Biodiscovery Ltd. This technique offers a unique solution for analyses of nanoparticles by utilising automated single particle trapping with Raman spectroscopy. This means SPARTA® can provide high-throughput biochemical information, in the form of Raman spectral fingerprints of individual nanoparticles, enabling us to distinguish between nanoparticles within heterogeneous mixtures, which would not be possible with bulk techniques. Research in the Stevens group has indicated a tremendous potential for sensitive and specific detection of disease vs. healthy cell derived nanoparticles, such as extracellular vesicles, using SPARTA®.

Alexandra Hopkins is a PhD student in Professor Dame Molly Stevens’ group at the University of Oxford, where her research focuses on harnessing Raman spectroscopy for the early diagnosis of Parkinson’s disease. Working alongside the spinout company SPARTA® Biodiscovery Ltd, this research sits at the intersection of physics, chemistry and biology, with particular relevance for minimally invasive, next generation diagnostics. Prior to her doctoral studies, Alexandra completed both her MSc and BSc in Biomedical Engineering and Biochemistry at Imperial College London.

Big Molecules and Small Particles

Prof. Lennart Lindfors, Pharmaceutical Sciences, AstraZeneca, Sweden.

Modified messenger RNA constitutes an interesting new approach for transient protein expression in different therapies. However, the details of the intracellular delivery of such macromolecules using lipid nanoparticles remains unknown. In this work we have prepared lipid nanoparticles (LNPs) of two different ionizable lipids (DLin-MC3-DMA and DLin-DMA), cholesterol, distearylphophatidyl choline (DSPC) and a PEG lipid. We then dosed LNPs intravenously in mice measuring LNP uptake, mRNA delivery and the concurrent protein expression in liver cells, i.e. hepatocytes, liver sinusoidal endothelial cells (LSEC) and Kupffer cells (KC). The in vivo data clearly showed that although uptake of lipid and delivered mRNA is very similar for both types of LNPs, the protein expression in hepatocytes is order of magnitude different.

In order to rationalize these in vivo observations, mRNA LNPs were characterized by several techniques e.g. 13C-NMR and small-angle x-ray scattering. Previously, we have shown that LNPs have a core-shell structure and here we focused our efforts into studying the core of LNPs, as bulk phases. The experimental efforts are backed up by a combination of all-atom, coarse-graining simulations and theory. By careful analysis of the inverse hexagonal phase structure of the ionizable lipids, we put forward a hypothesis on why DLin-MC3-DMA LNPs outperforms DLin-DMA LNPs in vivo.

Lennart Lindfors is currently a Senior Principal Scientist at Pharmaceutical Sciences at AstraZeneca with a PhD in Physical Chemistry at Chalmers University of Technology (1988). At AstraZeneca he has been working on oral vaccines, molecular simulations, nanoparticle systems of poorly soluble drugs and, recently, on lipid nanoparticles and exosomes incorporating messenger RNA.

Multiple Process Analytical Technologies for Optimisation of Freeze Drying – Digital Lyo

Honorary Professor Paul Matejtschuk, Department of Health and Life Sciences, De Montfort University, Leicester, UK.

Freeze drying is a key technology for delivering stable biopharmaceutics. Optimisation of the freeze drying cycle is of crucial importance in delivering economic and cost effective processes. As such the process engineering and underlying physical properties are critical to such optimisation. The use of existing Process Analytical Technologies (PAT) along with more novel technologies such as Through Vial Impedance Spectroscopy (TVIS) and Raman Laser spectrometry were investigated within the grant collaborative team with a model biotherapeutic formulation and common model proteins (IgG , BSA) to investigate the utility of these methods and to reduce primary drying times while maintaining product stability.

Paul Matejtschuk currently collaborates with Prof Geoff Smith who leads the DMU Lyo group. He has also served as Associate Lecturer at University of Surrey Department of Chemistry and Chemical Engineering, delivering postgraduate training in lyophilization. Previously Paul headed the Formulation Science Section at the Medicines and Healthcare products Regulatory Agency, at their South Mimms laboratories, in the UK, and over his career has developed freeze drying protocols for a wide range of biologicals .

Paul gained a BSc in Biochemistry from University of York (graduated 1982) and completed a PhD in Protein Chemistry at University of Warwick (awarded 1986) and has 40 years postdoctoral experience. He is a Director of the International Society for Lyophilization/Freeze Drying (www.islyophilization.org) and a committee member of the Thermal Methods Group of the Royal Society of Chemistry (www.thermalmethodsgroup.org.uk). He has successfully co-supervised five PhD studentships, with UCL and Imperial College London, published 60 peer-reviewed papers in addition to trade journal articles and book chapters and lectured widely in the field. He co-edited a book on freeze drying with Dr Kevin Ward (Lyophilization of Pharmaceuticals and Biologicals, New Technologies & Approaches, Ward KR & Matejtschuk P, Springer Press, 2019). Paul Matejtschuk (0000-0002-4646-800X) - ORCID

Label-free tracking of nanomedicines and small molecules in tissues with coherent Raman imaging

Prof. Julian Moger, Department of Physics and Astronomy, University of Exeter, UK.

Understanding how a bioactive small molecule penetrates biological barriers, distributes to its target tissue, and engages its molecular target is a fundamental challenge in both pharmaceutical and agrochemical science. Direct, spatially resolved imaging of unlabelled molecules in intact biological tissue remains technically demanding, and methods that provide distribution maps and mechanistic information simultaneously are scarce.

This talk presents label-free imaging methods developed at the EPSRC CONTRAST facility, the UK's dedicated broadband coherent anti-Stokes Raman scattering (BCARS) microscopy facility, validated in the context of agrochemical science and with direct translational relevance to pharmaceutical research. Using BCARS combined with machine learning-driven spectral analysis, we map the uptake and translocation of systemic fungicides in wheat leaves at subcellular resolution, without chemical modification of the compound or extrinsic labelling. We extend this to explore fungicide modes of action in pathogens, providing a spectroscopic readout of target engagement rather than a proxy pharmacodynamic measurement.

The physical challenges of imaging small molecule distribution in plant tissues are directly analogous to those in skin penetration studies, oral drug absorption, and tissue pharmacokinetics: heterogeneous matrices, low analyte concentrations, and spectrally complex biological backgrounds. These agrochemical case studies demonstrate a label-free small molecule imaging platform that is ready to be tested in pharmaceutical contexts.

Julian Moger is Professor of Biophotonics in the Department of Physics and Astronomy at the University of Exeter. His research focuses on the development and application of coherent Raman scattering (CRS) microscopy for label-free chemical imaging of biological systems. He established the EPSRC CONTRAST facility, the UK's dedicated broadband CRS user-access facility, which supports research across cancer diagnostics, pharmaceutical science, and agrochemical imaging. His group has applied

CRS to problems ranging from nanomedicine uptake and dermal drug delivery, to small molecule distribution in plants and fungal pathogens.

Optical coherence tomography velocimetry for the in-line processing of biologics.

Dr. Owen Watts Moore, Biological Physics, Department of Physics and Astronomy, University of Manchester, UK.

The flow of complex fluids is ubiquitous across a huge range of industrial processes, not least in the production of pharmaceuticals. For safety and consumer satisfaction, it is important that these products are formulated with specific physical properties, a task often complicated by variation in the base components. The most common solution is to take samples at key points in the formulation process for ex situ analysis, consuming energy, length ening production timescales, and generating significant waste due to the disposal of faulty batches. This makes an in-line method for quality control an attractive prospect for improving production line efficiency, helping to reduce costs and the environmental impact. Optical coherence tomography velocimetry (OCT-V) has recently emerged as a tool for studying the rheology of opaque complex fluids, allowing spatially and temporally resolved velocity measurements.[1–6] OCT-V uses the coherence length of its light source to section samples, suppressing multiple scattering artefacts that hinder other optical methods in opaque samples, thus making OCT-V a good candidate for in situ quality control. We now demonstrate the first OCT-V instrument for in-line processing, showing its ability to determine the physical properties of complex fluids, identify flow instability, and monitor the progress of mixing in situ in real manufacturing conditions on biologics and personal care products

Dr. Owen Watts Moore is a postdoctoral research associate in the biological physics group in the department of Physics and Astronomy at The University of Manchester. Working at the boundary of rheology and optical instrumentation, Owen’s primary research focus is developing optical velocity sensors for the in-line processing of complex fluids. The realisation of this work would allow in situ quality control on complex fluid production lines, reducing waste due to faulty batches, cutting energy demands, and saving time. After building a prototype apparatus in the lab, Owen and collaborators have successfully taken and published in-line measurements in real manufacturing conditions with antibodies, lamellar surfactant gels, and wormlike micelles.

Using atomic force microscopy to understand the bacterial cell wall architectural dynamics and its role in antimicrobial resistance

Dr. Abimbola Feyisara Olulana, School of Mathematical and Physical Sciences, University of Sheffield, UK.

The first part of my talk will focus on our recent publication on how MRSA survives in the presence of antibiotics. In Staphylococcus aureus, peptidoglycan (PG) is a 3D mesh-like macromolecule that surrounds the cell, playing an essential role in cell survival and stability by maintaining the shape of the cell during division and protecting the cell against its internal turgor pressure. Its biosynthesis proteins known as penicillin-binding proteins (PBPs) are the targets of ß-lactams antibiotics. In our previous works, we have shown the detailed molecular architecture of methicillin-sensitive Staphylococcus aureus (MSSA) peptidoglycan and the nanometric changes on the PG when treated with antibiotics(1, 2). In this study, we focused on the peptidoglycan associated with the methicillinresistant S. aureus (MRSA) and explore the following questions: 1) Are there differences in PG architecture when MRSA cell is treated with methicillin? 2) Can we genetically switch ON and OFF these nanoscale architectural phenotypes without using antibiotics? 3) Are these differences obtainable in clinical strains? I will speak on how we employed high resolution atomic force microscopy to image with nanometric resolution different surfaces (inner surface of the septum, outer surface of the septum, and outer surface of the cell periphery) of the extracted purified peptidoglycan from different derivatives of MRSA. Qualitatively, our results show distinct nanoscopic phenotype induced by antibiotic-treatment on resistant strains (both lab and clinical strains), displaying an alternative type of PG architecture at the site of cell division(3). Specifically, in the absence of antibiotics, the outer surface of the septum is characterized with concentric-ring architecture while in the presence of antibiotics the concentric ring is replaced with disordered dense mesh PG architecture(3). In addition to the nanoscale qualitative images that we obtained from the AFM, we also developed two analysis pipelines; One for PG fibre orientation detection and the other for quantitative measurement of the pore area that are associated with different surfaces and strains that we examined in our study.

The second part of my talk centres on the manuscript that we are currently working on, which has to do with the E.coli peptidoglycan (PG) architectural dynamics when treated with mecillinam. Unlike S.aureus, E.coli is a rod-shaped Gram-negative bacterium whose shape is maintained by 2D periplasm-based biopolymer known as Peptidoglycan (PG)(4). The chemical composition of PG is well understood(4) but the following questions remain unanswered; 1) what is the detail molecular organization of the PG; 2) is its organization location-dependent or not; 3) In the case of antibiotic induced shape change and death, what happens to the PG organization? To this end, we used high resolution atomic force microscopy (AFM) to track and map the changes in the PG organization from the pole to the cylindrical section of the rod. We extend this location-dependent imaging to interrogate different areas of the PG under different antibiotic treatment times. I will talk about the remarkable nanoscale architecture that we observed as a function of location on the PG and the exposure time with antibiotics. The aforementioned analysis pipelines were used in this study and we are able to link nanoscopic phenotypes like orientation to the specific activity of PG machinery that makes such ordered structure and how the bigger holes in the PG networks lead to instability of the PG that precedes death of E.coli cells. These two studies show the versatile application of AFM in bringing to surface new understanding about the bacterial cell wall and its role in life, death, and resistance

Dr. Abimbola Feyisara Olulana is a research associate in Prof. Jamie Hobbs Lab at the School of Mathematical and Physical Sciences, University of Sheffield. Her work focusses on utilising high resolution atomic force microscope to profile, investigate, and quantify the inherent nanoscale structural motifs and topographical properties associated with peptidoglycan harvested at the exponential phase of different methicillin-resistant Staphylococcus Aureus (MRSA) and the Escherichia coli (E.coli) derivatives. Here are some of the research questions that her work seeks to address; 1) How does resistant changes the cell wall architecture? 2) Can we distinguish the antimicrobial strains based on the material properties of their associated cell wall? And 3) at the nanoscale what happens to cell wall architecture when treated with antibiotics? Her work promises to yield more understanding as to how antimicrobial resistant impacts on the cell wall architecture and mechanics, leading to promising designs for antibiotics that can circumvent resistant in bacteria.

Microscopic cluster formation and viscosity in monoclonal antibody solutions - drug formulation meets colloid physics

Dr. Tilo Seydel, Institut Max von Laue – Paul Langevin, Grenoble, France.

Monoclonal antibodies (mAbs) have gained importance for therapeutics due to their specificity and versatility. Self-administration by subcutaneous injection reduces the number of hospital visits, but requires antibody solutions that are highly concentrated at a limited maximum viscosity for injection. In this context, crowding and protein–protein interactions (PPIs) challenge the stability, manufacturability, and delivery of a mAb drug. The understanding of macroscopic viscosity requires knowledge of protein diffusion, PPIs, as well as of self-association or aggregation. We present a systematic diffusion study of a series of different mAbs of the IgG1 subtype in aqueous solution depending on the concentration and temperature by quasi-elastic neutron scattering (QENS). QENS probes the short-time self-diffusion of the molecules and therefore accesses the hydrodynamic mAb cluster size and the superimposed internal mAb dynamics. Small-angle neutron scattering (SANS) is jointly employed to probe structural details and to understand the nature and intensity of PPIs. Complementary information is provided by neutron spin-echo spectroscopy (NSE), molecular dynamics (MD) simulations and viscometry, thus obtaining a comprehensive picture of mAb diffusion.

Tilo Seydel graduated with a PhD in physics from the University of Kiel in northern Germany in 2000, having employed X-ray reflectivity and photon correlation spectroscopy using synchrotron radiation to study lateral surface structure and dynamics at the glass transition. He subsequently moved to the Institut Max von Laue - Paul Langevin (ILL) in Grenoble, France, where he has contributed to the development of neutron optics for high-resolution spectroscopy, including the phase space transformation technique, and simultaneously established a research program on biologically inspired soft matter. Being an ILL scientist, he also holds an appointment with the University of Grenoble-Alpes (UGA) doctoral school for the supervision of PhD projects, following a habilitation degree. In collaboration with the UGA and partner universities in ILL member countries, he has supervised and co-supervised numerous Master, PhD, and postdoctoral research projects. His recent research involves external grants as well as partnerships with pharmaceutical companies.