Alain Pluen, PhD., B.Sc., M.R.S.C.

DPO Lead for Post-Graduate Teaching

Post-Graduate research Tutor

Alain Pluen – a physical chemist by training – is a Senior Lecturer within the Division of Pharmacy and Optometrics. Alain joined the then School of Pharmacy in 2000 and has been pursuing his research effort on studying protein-protein interactions espectially aggregation in solution to understand issues related to biologics (e.g. mAbs) formulation and cells.  This is carried out using bioimaging techniques (confocal microscopy, Raster Image Correlation Spectroscopy or Fluorescence Correlation Spectroscopy) as well as light scattering (DLS, SLS).

A second area of research focuses on nanoparticles especially their formulation. In particular RNA lipid nanoparticles in order to extent the shelf life of these and reduce the reliance on the cold chain, and, hyrophobic ion pairs to improve the delivery of antimicrobials.

 Previously Alain did his post-graduate studies at the Institut Charles Sadron – Centre de Recherche sur les Macromolecules, (CNRS UPR 22, Strasbourg, France) where he studied the mechanisms of transport of macromolecules (single stranded DNA) in gels to improve sequencing (EU Biomed 1). Then in 1997, he joined the team of Prof. R.K. Jain at the Massachusetts General Hospital - Harvard Medical School as a post-doctoral research fellow. In this biomedical engineering laboratory and in collaboration with Dr. David Berk, he started to study the relation between the tumour microenvironment and the transport hindrance of novel medicines i.e. antibodies, protein, peptides or liposomes. In September 2000, he joined the then School of Pharmacy and Pharamceutical Sciences.

Member of the Royal Society of Chemistry

Fellow of the Higher Education Academy

Member of the Academy of Pharmaceutical Sciences

• 1996 Ph.D. Polymer Physical Chemistry - University of Strasbourg, France
• 1992 M.Sc. Physical Chemistry - University of Strasbourg, France
• 1991 B.Sc. Physical Chemistry - University of Strasbourg, France

UG teaching

• Medicine modules: PHAR11001, PHAR11002, PHAR33001
• Block lead PHAR22002

PG teaching

• PIAT: module 71010 (tutor), dissertation board and examination board

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. Improve the formulation of nanoparticles in particular RNA loaded lipid nanoparticles (effect of sterol variants on the stability of particles over time and different storage conditions) or the formation of hydrophobic ion pairs nanoparticles to deliver antimicrobial peptides.

3. 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 mechanisms of protein aggregation using ManICS and Spatial Intensity Distribution. These concepts have also been applied to transport in biofilms (Rattray et al. Methods in Molecular Biology 2019, Buzza et al., Microbiology spectrum, 2023)

4. Exfoliation and stabilisation of pristine graphene suspensions using short peptides (Alkathiri et al., J Phys Chem C 2019)

5. Understanding the delivery of drugs or biologics 

• Cell penetrating peptides e.g. Tat (Saleh et al., Biopolymers 2008) and amphipathic, DNA-binding cationic peptides e.g. LK15 (Perry et al., J. Fluorescence 2008). This lead to the design of the multifunctional peptide, TatLK15, which successfully mediates cell transfection in cell monolayers and spheroids (Saleh et al., J Controlled Release 2010) andefficiently mediated the silencing of bcr-abl oncoprotein using siRNA and shRNA (Arthanari et al., J controlled Release 2010). We are currently investigating the behaviour of the peptide itself, its pro-apoptotic acitvity and its ability to overcome multidrug resistance. We are currently refining this multifunctional peptide to improve its efficiency in primary cells.

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).

Collaborators

Manchester (UK)

Divivsion of Pharmacy and Optometry

Dr H Aojula

Dr Richard Bryce

Prof A McBain

Manchester Integrated Biosciences

• Ms Lauren Daniels, PhD student, CTP with Astra Zenecca
• Mr Mohammed Alosaimi
• Mr Mark O'Hanlon

Alumni:

• Dr Ibrahim Alsasabih, former PhD student
• Dr James Austerberry, former PDRA, Ipsen
• Dr Fai Alkhathiri, former PhD student, Assistant Prof, King Saud University
• Dr Maria-Laura Grecco, former PhD student,Astra Zenecca
• Dr Nicole Sibanda, former PhD student, Kymab ltd
• Dr Catriona McCallion, former PhD student, civil servant
• Dr Ben Staley, former PhD student, Accor UK.
• Dr Peter Davis, former PDRA, Albumedix.
• Dr Myasar Al-kotaji (former PhD student), visiting researcher UCL, Head of Pharmaceutics at Mosul University (Iraq).
• Dr Zahra Hamrang, former PhD student and PDRA, Sr Lecturer, School of Pharmacy, Strathclyde University.
• Dr Yamini Arthanari, former PGR student,Oxford Biomedica.
• Dr Maryam Shah, former PGR student, Proveca Ltd
• Dr Heather Perry. Heather, former PGR student, Waters corp.
• Dr Sylvie Lélu, former PGR student and post-doc, Institute of Physics National Technological University of Norway
• Dr Hayley McGlynn, former PGR student, Novartis.
• Dr Ning Ning, former PG  student, Astra Zenecca.
• Dr Cyril Rauch, former PDRA, Associate Prof, School of Veterinary Sciences, U. of Nottingham, UK
• Dr Amer Saleh, former PGR student, Astra Zenecca.
• Mr Xiaofei Lei (Hank), former Master student.

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

Models:

• 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

Alain Pluen takes regularly Nuffield placement students during Summer time and is also a school governor of a primary school in the Peak District