COEBP Biopharmaceutical Research Feb2014

Biopharmaceutical Research

University of ManchesterCentre of Excellence in Biopharmaceuticals

www.coebp.ls.manchester.ac.uk

mailto:[email protected]

Centre of Excellence in Biopharmaceuticals Biopharmaceutical Research

This presentation lists a number of research projects which are potentially relevant to companies that discover, develop or manufacture protein biopharmaceuticals.

Some are projects already underway or completed. Others are ideas which require further industry input before seeking funding.

The Centre’s Research is grouped into two themes underpinning industry research:

Product characterisation and formulation

Cell line development

University of Manchester Biopharmaceutical Technologies

Mechanisms of Protein Aggregation

Dr Robin [email protected] +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




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” [email protected] +44161 306 5813




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 [email protected] +44161 306 4887




Applications of RICS to Formulation and Delivery of Biologics

• 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 [email protected] +44 (0)161 275 1792




PK/PD Modelling for Biologics

• Adapt existing models for small molecule PKPD simulations

• Simulate different technologies eg – Half life extension (PEGylation,

fusion proteins)– Bispecifics– Antibody Drug Conjugates

• IV, subcutaneous, oral, inhaled biologics

• Tissue penetration and microscopic distribution

• Potential use in improved biologics molecular design, dosage, trial design

Dr David [email protected] +44 (0) 161 275 2375




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 [email protected] 0161-306 4490



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 [email protected] +44161 306 4207

Prof Ian [email protected] +44161 275 1587




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 [email protected] +44161 306 4480




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 [email protected] Phone +44161 306 5819



NMR Investigations of Protein Structure and Dynamics

Prof Jon [email protected] +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



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 [email protected] +44161 306 4537



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 [email protected] +441612755317




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 [email protected] +441612755317

Dr Graham [email protected] +441613064477






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

[email protected] +441612751554




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 [email protected] +441612755077




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 [email protected] +441612755462

Prof Mark [email protected]+44 (0)161 275 3921





Protein Biochemistry & Expression

Prof. Tony [email protected] +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



If you are interested in any of these projects

please contact

University of ManchesterCentre of Excellence in Biopharmaceuticals

[email protected]@manchester.ac.uk

[email protected]

www.coebp.ls.manchester.ac.uk


COEBP Biopharmaceutical Research Feb2014

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