Alain Pluen’s research team focuses on:
1. Understanding weak protein protein interactions especially for biopharmaceuticals using fluorescence techniques, light scattering.
In collaboration with Drs Curtis, Warwicker, Golovanov Profs Derrick and Dickson (MIB and FBMH). This work is supported by BBSRC and MedImmune. Aggregation propensity of mABs in simple solutions as well as real formulations has been deternined and compared to standard equipment (Hamrang et al. 2015 JPharm Sci and Shah et al. 2017 Int J Pharm). The effect of charge mutations on teh stability and aggregation of small chain Fv fragments studied (Austerberry et al., Eur J Pharm and Biopharm).
2. Develop the use of Image Correlation Spectroscopies for quantifying :
- peptides uptake in cells (Staley et al., Drug Discovery Today 2010) and to
- protein and biologics aggregation phenomena in collaboration with the Centre of Excellence in Biopharmaceuticals (COEBP).
- We have demonstrated the ability of Raster Image Correlation Spectroscopy at characterising the diffusional behaviour of proteins in solution under normal and stress conditions (Hamrang et al.,J Pharm Sci, 2010): using diffusion coefficients and local concentrations, variations of the particles distribution illustrate the effect of stress on protein solutions. A collaboration with Dr R Curtis (MIB) focuses on te mechanismss of protein aggregation using ManICS and Spatial Intensity Distribution Analysis among other techniques. Another element of this approach is the characterisation of protein aggregation on cells surfaces with Dr P Legembre (Rennes France).
3. Preparation of stable graphene suspensions and their for drug delivery.
4. Understanding the delivery of drugs or biologics
Additionally, in collaboration with Dr Rauch (U. of Nottingham) the influence of phospholipids e.g. phosphatidylserine on drugs uptake in tumour cells is investigated (Rauch and Pluen, Eur J Biophysics 2007) as well as multidrug resistance (Rauch et al., Cellular Biochemistry and Biophysics 2010).
Fluorescence techniques (Zeiss LSM510 Meta Confocor 2):
- confocal laser scanning microscopy cells
- Image correlation spectroscopy i.e. TICS, RICS - Designed our own software (ManICS).
- fluorescence correlation spectroscopy (FCS) in solutions and cells
- fluorescence recovery after photobleaching (FRAP) in solutions, gels, cells
- cell monolayers (tumours cells)
- tight monolayers (Caco-2, endothelial cells)
- multicellular tumour spheroids
- flow chamber
- composite gels
Peptide synthesis in collaboration with Dr H Aojula