ERC Advanced Grant (ERC grant 227950): "Enhancing microfabricated devices with chemical imaging for novel chemical technology" (PI: S. G. Kazarian)

In this research, in collaboration with the Faculty of Medicine at Imperial College London, we demonstrate in situ chemical imaging approach to study in situ protein crystallisation by macro ATR-FTIR imaging. This work presents an important advance since it allowed us to monitor chemical and physical changes in situ with fast acquisition time, satisfactory spatial resolution and high chemical specificity provided by FTIR imaging. This methodology could provide a convenient method for validation of crystallising proteins in situ to differentiate real protein crystals from other crystals (e.g. salt) or pseudo-crystals. This in situ imaging method can be used as a tool to screen multiple samples in high-throughput manner, thus optimising the condition for growing large monolith protein crystals. This research was described in the cover article in Analytical Chemistry (2009), which followed by article on protein aggregation  in Analytical Chemistry (2014) and by most recent studies on protein purification (ABC 2015 and Scientific Reports 2016), these latest studies have been the result of a successful joint BBSRC project with Dr. Bernadette Byrne (Department of LIfe Sciences at Imperial College London) with whom we previously jointly supervised a PhD student Stefanie Glassford on applications of FTIR spectroscopic imaging to studies of proteins. We now continue our collaboration via a  PhD student Hannah Tiernan on project "Conformational Changes and Agglomeration in Biopharmaceuticals Using Spectroscopic Imaging in Microfluidics" which started in October 2017. We also collaborate via Mr. James Beattie on his PhD project “Monitoring of protein purification by spectroscopic techniques” which involved collaborfation with GSK. Our most recent article on ATR-FTIR Spectroscopic Imaging for the Analysis of Biopharmaceuticals (doi) has been recently featured in GEN (Genetic Engineering & Biotechnology News).   

Key References

• Chan K. L. A., Govada L., Bill, R. M., Chayen, N. E., Kazarian S. G., ATR-FTIR spectroscopic imaging of protein crystallization, Analytical Chemistry 81 (2009) 3769-3775. (COVER article above) This article is openly available to view from here   (doi)
• Glassford S., Chan K. L. A., Byrne B., Kazarian S. G. Chemical imaging of protein adsorption and crystallisation on a wettability gradient surface Langmuir 28 (2012) 3174-3179 (doi)
• Glassford S., Govada L, Cheyen N. E., Byrne B., Kazarian S. G. Micro ATR FTIR imaging of hanging drop protein crystallisation Vibrational Spectroscopy 63 (2012) 492-498 (doi)
• Glassford S. E. , Byrne B., Kazarian S. G. Recent applications of ATR FTIR spectroscopy and imaging to proteins BBA – Proteins and Proteomics  (2013) 1834, 2849-2858(doi) (Free Access)
• Boulet-Audet, M., Byrne, B., Kazarian, S. G. High-throughput thermal stability analysis of a monoclonal antibody by ATR-FTIR spectroscopic imaging Analytical Chemistry (2014) 86(19), 9786–9793. (doi)  (Open Access article)
• Boulet-Audet M., Byrne B., Kazarian S. G.  Cleaning-in-place for immunoaffinity resin monitored by in situ ATR-FTIR spectroscopy  Analytical and Bioanalytical Chemistry (2015) 407, 7111-7122. (doi - Open Access)
• Boulet-Audet M., Byrne B., Kazarian S. G. In-column ATR-FTIR spectroscopy to monitor affinity chromatography purification of monoclonal antibodies  Scientific Reports (2016) 6, 30526  (doi)
• Tiernan H., Byrne B., Kazarian S. G. Insight into heterogeneous distribution of protein aggregates at surface layer using ATR - FTIR spectroscopic imaging. Analytical Chemistry (2020) 92 (7) 4760-4764 (doi)
• Tiernan H., Byrne B., Kazarian S. G.  ATR-FTIR spectroscopy and spectroscopic imaging for the analysis of biopharmaceuticals.   Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy (2020)  241, 118636 (doi) has been featured in GEN (Genetic Engineering & Biotechnology News).