1. Biopharmaceutical Research
University of Manchester
Centre of Excellence in Biopharmaceuticals
www.coebp.ls.manchester.ac.uk

2. Centre of Excellence in Biopharmaceuticals
Biopharmaceutical Research Project
List
Mechanisms of Protein Aggregation – Dr Robin Curtis
Concentrated Protein Solutions – Dr Sasha Golovanov
Protein Solution Rheology – Dr Xue-feng Yuan
Raster Image Correlation Spectroscopy – Dr Alain Pluen
Predictive modelling of Protein Solubility – Dr Jim Warwicker
Immunogenicity of Protein Aggregates and SVPs – Profs Jeremy
Derrick & Ian Kimber
Detection of Protein Contaminants in Biopharmaceuticals – Prof
Roy Goodacre
Higher Order Structure of proteins – Dr Ewan Blanch
RiboTite - Controllable Protein Expression in bacteria – Dr Neil
Dixon
CHO cell line development – Prof Alan Dickson
Differentiation of Single Stem Cells – Prof Mark Dunne, Dr Karen
Cosgrove
Molecular organisation and function of glycoprotein polymer gels –
Prof Dave Thornton

3. University of Manchester Biopharmaceutical Technologies
Mechanisms of Protein Aggregation
Dr Robin Curtis
r.curtis@manchester.ac.uk
Phone +441613064401
• Develop improved methods for
predicting and controlling aggregation
during processing and in liquid
formulations using a combination of
numerical and experimental studies
• Delineate effects of partial unfolding
and protein self association on
aggregation kinetics
• Partial unfolding probed with
intrinsic/extrinsic fluorescence. Static
and dynamic light scattering used for
probing protein-protein interactions in
terms of osmotic second virial
coefficient, B22
• Examine link between protein-protein
interactions and protein phase
separation/opalescence

4. University of Manchester Biopharmaceutical Technologies
Concentrated Protein Solutions
• Method for solubilisation of
proteins, with increased stability
during bioprocessing and storage
• 50mM L-Arg + L-Glu; solubility of
many proteins increased (up to 8
times)
• Further work in progress
• Golovanov AP, Hautbergue GM, Wilson
SA, Lian LY. (2004). A simple method for
improving protein solubility and long-term
stability. Journal of the American
Chemical Society, 126(29), 8933-9
Dr Alexander “Sasha” Golov
a.golovanov@manchester.a
Phone +44161 306 5813

5. University of Manchester Biopharmaceutical Technologies
Protein Solution Rheology
• Rheological characterisation of high
concentration protein solutions
• Modular design micro-rheometer
system
• Suitable for cost-effective mass
production
• High sensitivity
• Small sample required
• Potential for high throughput
Dr Xue-feng Yuan
Xue-
Feng.Yuan@manchester.ac
.uk
Phone +44161 306 4887

6. University of Manchester Biopharmaceutical Technologies
SVP, Aggregates Detection and
Counting
• Raster Image Correlation Spectroscopy
• Detects and counts protein
molecules, reversible self
aggregates, irreversible aggregates, sub-
visible particles in same sample
• Determines hydrodynamic size
• No need to dilute or filter sample
• Small sample
• Potential for high throughput
characterisation
• Raster image correlation spectroscopy as
a novel tool for the quantitative
assessment of protein diffusional
behaviour in solution, Hamrang Z, Pluen
A, Zindy E, Clarke D, Journal of
Pharmaceutical Sciences, 2012
Dr Alain Pluen
alain.pluen@manchester.ac
Phone +44 (0)161 275
1792

7. University of Manchester Biopharmaceutical Technologies
Predictive Modeling of Protein
Solubility
• The aim is to predict the solubility of
biopharmaceuticals based on
knowledge of primary sequence and
structure
• There are currently three approaches to
modelling protein solubility from
sequence and structure:
– partial unfolding
– beta-strand/amyloid-forming
propensity
– protein surface properties
• In preliminary studies, the best
correlation between surface properties
and solubility was for a mixture of non-
polar and polar features that takes into
account detailed surface geometry.
Dr Jim Warwicker
jim.warwicker@manchester.
0161-306 4490

8. University of Manchester Biopharmaceutical Technologies
Immunogenicity of Protein Aggregates
and Sub Visible Particles
• Aim to determine the
influence of aggregates of
well-defined reference
protein(s) on
• vigour of T lymphocyte
and IgG and IgE
antibody responses in
mice
• development of
functional
subpopulations on CD4+
T lymphocytes (Th1, Th2
and Treg cells) in mice
• phenotype and function
of dendritic antigen
presenting cells
Prof Jeremy Derrick
jeremy.derrick@mancheste
Phone +44161 306 4207
Prof Ian Kimber
ian.kimber@manchester.ac
Phone +44161 275 1587

9. University of Manchester Biopharmaceutical Technologies
Spectroscopic Detection of Protein
Contaminants
• Ability to detect protein
contaminants at low levels in
protein solutions using FT-
IR, Raman spectroscopy
• Apply chemometrics to the
comparison of spectroscopic data
• Preliminary data with FT-IR on
spiked proteins eg RNAse A spike
with 1% RNAse B
Prof Roy Goodacre
Roy.Goodacre@manchester.ac
Phone +44161 306 4480

10. University of Manchester Biopharmaceutical Technologies
Higher Order Structure of Proteins
• Characterize and quantify protein higher order
structure (secondary and tertiary) using
Raman and FT-IR spectroscopies (already 3
times more accurate than CD)
• Apply bioinformatics and chemometrics to
spectroscopic data to study structural changes
• Detection of flow-induced aggregation and
correlating protein structural changes with the
shear forces applied (with a colleague at
Mechanical Engineering, UCL)
• Structural characterization of glycoproteins
and carbohydrates
Dr Ewan Blanch
e.blanch@manchester.ac
Phone +44161 306 5819

11. University of Manchester Biopharmaceutical Technologies
NMR Investigations of Protein Structure
and Dynamics
Prof Jon Waltho
j.waltho@manchester.ac.uk
Phone +441613064191
• Structure, dynamics and energetics of
near transition-state complexes of
enzymes
• Characterisation of conformational
transitions within partially folded
states of proteins
• Intermediate species involved in the
assembly of prion particles, and the
inhibition of assembly processes by
small molecules
• Fundamentals of instability within
protein therapeutics

12. University of Manchester Biopharmaceutical Technologies
RiboTite Controllable Protein
Expression Technology
• Controls expression in bacteria
using synthetic compounds
• Regulates translation initiation
• Tight control prior to induction
• Affords true cellular level titratable
control
• Controlled co-expression of two or
more proteins
• Dixon N, Duncan JN, Geerlings
T, Dunstan MS, McCarthy JE, Leys
D, Micklefield J. Proc Natl Acad Sci U S
A. 2010 Feb 16;107(7):2830-5
Dr Neil Dixon
Neil.Dixon@manchester.a
Phone +44161 306 4537

13. University of Manchester Biopharmaceutical Technologies
Transcription in response to small molecules
• Protein production in yeast
• Structural biology of transcriptional complexes
• Understanding how small molecules and
metabolites can influence protein-protein
interactions to affect transcription
• Determining how changes in cellular location
of a protein can influence its function
• Protein expression and purification facility
Prof Richard Reece
Richard.Reece@manchester.
Phone +441612755317

14. University of Manchester Biopharmaceutical Technologies
Yeast expression technologies
• Transcriptional/translational control mechanisms
• Environmental signaling and stress responses
• Small molecule intervention with regulatory
processes
• Yeast genetics and physiology
• Yeast genetic engineering
Prof Richard Reece
Richard.Reece@manchester.ac
Phone +441612755317
Dr Graham Pavitt
Graham.Pavitt@manchester.ac.u
Phone +441613064477

15. University of Manchester Biopharmaceutical Technologies
Protein quality control in the ER
• Understanding the mechanisms of,
and regulation of protein folding
reactions in the ER and secretory
pathways
• Consequences for secreted protein
yield and quality
• Application of molecular and cellular
biology toolkit to dissect regulatory
networks
Dr Lisa Swanton
Lisa.Swanton@manchester.a
Phone +441612751554

16. University of Manchester Biopharmaceutical Technologies
CHO Cell Line Research
• Optimisation of CHO platforms via
„omics approaches, including
metabolic flux balance models and
rationalised feeding regimes
• CHO clonal variation analysis at
molecular and cellular level –
realtionship between phenotype and
clonal utilisation for defined product
portfolios
• CHO cell line stability at molecular
(genomic and wider cellular) level
• Early predictability of CHO cell lines in
relation to biomass
attainment, productivity and product
quality
Prof Alan Dickson
Alan.Dickson@manchester.a
Phone +441612755077

17. University of Manchester Biopharmaceutical Technologies
Differentiation of Single Stem Cells
• metabolic profiling studies
of a model stem cell
differentiation system
(pancreatic cells) at a
single cell level
• metabolic profile within
populations undergoing
differentiation
• follow metabolic profiles
from single cells during
differentiation
• to follow the metabolic
profile at single cell level
for cells that maintain an
undifferentiated phenotype
Dr Karen Cosgrove
Karen.Cosgrove@manchester.ac
Phone +441612755462
Prof Mark Dunne
Mark.J.Dunne@manch
ester.ac.uk
Phone+44 (0)161 275
3921

18. Molecular organisation and function
of glycoprotein polymer gels
• Structural and functional characterisation of
glycoprotein polymer networks – in vivo and vitro
• Glycoprotein purification/biochemistry
• Mass spectrometry
• Electron microscopy/AFM
• Microrheolology
• Biophysical analysis (MALLS/AUC/SAXS)
• Primary cells and cell lines
Professor Dave Thornton
Dave.thornton@manchester.ac.uk
Phone +44161 275 5647

19. University of Manchester Biopharmaceutical Technologies
Protein Biochemistry & Expression
Prof. Tony Day
anthony.day@manchester.ac.uk
Phone +44 161 2751495
• Protein biochemist with experience of
low and high resolution structural
analysis of proteins and protein-ligand
complexes
• Expertise in the analysis of protein-
ligand interactions by a wide range of
methodologies (Academic Lead for
BioMolecular Analysis Core Facility)
• Expression of proteins in E. coli. and
refolding/purification methodologies
• Ongoing development of protein
biological for the treatment of
osteoporosis and other disorders

20. If you are interested in any of these projects
please contact
University of Manchester
Centre of Excellence in Biopharmaceuticals
malcolm.rhodes@manchester.ac.uk
joanne.flannelly@manchester.ac.uk
alan.dickson@manchester.ac.uk
www.coebp.ls.manchester.ac.uk