Our third webinar in the MDC Connects Series 2021 | A Guide to Complex Medicines. This slide deck gives an overview of the early assessment of Prototype Nanomedicine Nano Bio Interactions. Zahra Rattray, University of Strathclyde

1. Dr Zahra Rattray
Chancellor’s Research Fellow
Laboratory of Complex Bioanalysis
MDC Connects Series 2021
May 2021
The Early Assessment of Prototype
Nanomedicine Nano-Bio Interactions
‘Where Nano meets Bio’
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zahra.rattray@strath.ac.uk

2. The Biological Fate of Complex Medicines
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Hard corona
Soft corona
Particle properties
Composition
Surface charge
Morphology
Size
Cargo
Concentration
…
Environmental parameters
Incubation media
Protein content and composition
Duration
Temperature
Flow rate

3. The Protein Corona: Why Measure the Protein Corona?
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Impact on drug release
Colloidal Stability
Cellular Interactions
(cytotoxicity, immunotoxicity, uptake)
• Cellular uptake
• Complement activation
• Macrophage activation
• Circulation time
• Relative organ distribution
Organ Biodistribution

4. 4
Multiscale Modelling of Nanomedicine Biological Fate
In
Silico

5. The Protein Corona Bioanalytical Toolbox- In vitro
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Media Nanoparticle
Co-incubation
Nanoparticle isolation/recovery
(ex situ/in situ)
Corona proteins/pelleted particles
Separation
Digestion (peptide fragments)
Mass
Size,
Concentration,
Structural
Circular Dichroism/FTIR
Raman
NMR
Quantification
Protein assays
LC-MS/ICP-MS
Imaging TEM/cryo-TEM
Identification
MS
SDS-PAGE
ELISA
Affinity/Dynamics
ITC/QCM
Reported for metallic, inorganic, polymeric, etc

6. In situ Profiling of the Protein Corona- Physical Analysis
Daramy et al, Unpublished data- Laboratory of Complex Bioanalysis
Mean particle dry mass change relative to PLGA
nanoparticles treated in 10% FBS (n=3). * represents a
P<0.05, ** P<0.01, *** P<0.001. 6
Example- PLGA Nanoparticles
Resonant Mass Measurement PTA RMM

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The Protein Corona Bioanalytical Toolbox
Alberg, I., et al. (2020). Small 16(18): 1907574.
Weber, C., et al. (2018). Acta Biomaterialia 76: 217-224.
Weber, C., et al. (2019). Biomacromolecules 20(8): 2989-2999.
Field Flow Fractionation Doxorubicin liposomes in cell culture
media- EAF4 with light scattering
Other hyphenations of FFF exist!
Strathclyde Multiscale Metrology Facility (Q4 2021)
Field Flow Fractionation (asymmetric, electric and centrifugal) with
multiple light scattering, Raman and ICP-MS detection
Drexel et al. (2020). Molecules 25(20), 4703.
Asymmetric FFF
Centrifugal (CF3)
Electrical (EAF4)
SEC

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The Protein Corona Bioanalytical Toolbox- In vitro
Jayaram D.T., et al. (2018). Biophys J 114: 209-216.
Static versus flow conditions
Example- Polystyrene model nanoparticles (200 nm)
•Flow impacts corona composition
•Protein conformation changes between static vs flow
•Protein-dependent effects
•This effect has been observed with lipid nanoparticles

9. In the era of precision medicine …
• Microarray based assays for mimicking biorelevant conditions
• Multiplexing analytical measurements
• Multiscale models predicting and measuring the protein corona  prototype
Line of Sight- The Precision Protein Corona Interactome
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10. • Parallel physical and compositional assessment
• Biorelevant conditions
• In situ versus ex situ approaches
• Harmonization of reporting for biological interactions
• Interactome passport for each prototype!
• Combining in silico output with in vitro fingerprint
Summary
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11. AstraZeneca
Past colleagues, too many to name!
Laboratory of the Government Chemist (LGC)
Dr Heidi Goenaga-Infante
Medicines Discovery Catapult
Dr Martin Main, Dr Graeme Wilkinson, Dr Duygu Yilmaz
Malvern Panalytical
Elise Guerini, Liam Cole
National Physical Laboratory
Caterina Minelli, Yiwen Pei
University of Strathclyde
Prof Duncan Graham (Pure & Applied Chemistry)
Prof Karen Faulds (Pure & Applied Chemistry)
Prof Yvonne Perrie (Pharmacy & Biomedical Sci)
Dr Nicholas Rattray (Pharmacy & Biomedical Sci)
Dr Asimina Kazakidi (Biomedical Engineering)
Prof Gavin Halbert (Pharmacy & Biomedical Sci)
Yale University
Dr James Hansen, Dr Caroline Johnson, Dr Jiangbing Zhou
Acknowledgments
Laboratory of Complex Bioanalysis
Miss Layla Alnoumas
Mr Abdullah Alsultan
Miss Zoe Apczynski
Mr Bhushan Bhale
Miss Domenica Berardi
Mr Karim Daramy
Mr Ahmed Gaafar
Ms Merrit Rothe
Mx Kiri Thornalley
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