“Miniaturisation and automation of bioprocess development
holds great promise in reducing the time and cost to market of new
biopharmaceuticals. This meeting aims to highlight recent technologies used in
high throughput bioprocess development, from clone selection through to
analysis of final product and formulation. A series of expert speakers will
describe the development and use of current miniaturisation technologies
together with the technical and regulatory hurdles that must be overcome to
facilitate wider industrial uptake."
This event has CPD accreditation and will have a
discussion panel session.
On registration you will be able to submit your questions
to the panel that will be asked by the chair on the day of the event
Meeting Chair - Professor Gary Lye, Professor of Biochemical
Engineering, University College London
8:45 – 9:30
Registration
9:30 – 9:45 Introduction
by the Chair: Professor Gary Lye, Professor of Biochemical
Engineering, University College London
9:45 – 10:10
Scale Down Approaches to Facilitate CHO Clone
Development for High-Level mAB Expression
Dr Ray Field,
Director Cell Sciences, MedImmune, Cambridge, UK.
With increasingly diverse portfolios of biotherapeutics,
more efficient methods for creating and screening high expressing stable cell
clones are starting to be used. The ability to screen multiiple
clones expressing multiple recombinant protein and mAB candidates for each
therapeutic project is becoming a requirement in order to identify the
appropriate clone / mAB combination for effective drug development. Increasingly,
shaking microwell and microbioreactor scale down models are being used to
facilitate such clone selections leading to increased predictibility of
Bioprocessing. Strategies, Issues and case studies will be described
to illustrate this approach.
10:10 – 10:35 High Throughput Process Development
Technologies: Successful Implementation and Application to Process
Improvement.
Dr Jonathan Dempsey, Life Technologies, UK
Biopharmaceuticals are an increasingly important class of
medicines both in terms of meeting unmet medical needs and commercially. These
molecules are manufactured using living organisms and by necessity the
processes used are complex, time consuming and costly. In order to increase the
availability and reduce the cost of biological medicines novel methods are
needed to improve the development phase of these products Automation of
biopharmaceutical development holds great promise in increasing the quantity of
these molecules which can be produced and reducing the time and cost to market
of these medicines. In this presentation I describe the implementation of two
automated cell process development systems, the benefits and limitations of
their use and data demonstrating increased throughput and enhanced processes
10:35 – 11:00 The development of a 24/48 vessel automated
micro-scale bioreactor and comparison to bench-scale bioreactors
Dr Kenneth Lee, The Automation
Partnership, UK
Industrial antibody production and many other commercial
bioprocesses rely on the selection of the final industrial cell line from
hundreds of clones. With such a large number of clones to screen it is
inevitable that a large proportion of the clones, possibly over 95%, will not
make it to the bioreactor stage: the point at which bioprocesses can be
adequately modelled and scale-up principles can be applied. The ideal selection
process would be to run a stability check on the system, followed immediately
by bioreactor trials. However, the setup and running of bioreactors is
laborious, time consuming, and costly. The result is that only the top 1 – 2%
of clones are ever evaluated in bioreactors, usually without replication.
Static well plates and shake flasks, often used to bridge the gap between stability
study and the final clone selection, are inherently different to the
bioreactor: static plates have no mechanical input and shake-flasks only have
passive control of dO and pH, so high performing clones that have a small
process window may be discarded.
Bioreactor mimics enabling highly parallel culture and minimising the
requirement for labour, provide a solution to this problem. Additionally lower
volume systems with reduced footprint decrease costs and minimise the impact on
valuable laboratory space. Results from
trials of a novel 24-vessel automated micro-scale bioreactor mimic (ambr™) showed very comparable results compared to
5-litre and 10-litre bioreactors.
Experiments investigating the growth characteristics in AMBR are very
favourable: batch growth shows very tight coefficient of variation (CV) on cell
number and viability. Antibody production in fed-batch trials also showed very
good comparability with 5-litre and 10-litre bioreactors: a 10 cell clone trial
in bioreactors and AMBR showed very similar growth curve profiles, final cell
number, and ranking in cell clone productivity. In addition the values for cell
productivity rate were also very similar between AMBR and bioreactor.
11:00 – 11:05 Speakers
photo
11:05 – 11:30
Mid-morning break
11:30 – 11:55
Microscale
to Manufacture for Emerging Vaccine Technologies
There has been a recent upsurge in the interest in using viral vectors such as
adenoviruses, in the field of viral vectored vaccines and gene therapy. It is
estimated that there are currently 377 clinical trials world wide conducted in
association with this class of therapeutic product. The recent increase in demand,
coupled with the relatively high titres needed for pre-clinical and clinical
trials has fuelled the requirement for a new approach to adenoviral
production. This presentation describes Eden Biodesign’s innovative
approach to the rapid development and implementation of a robust, scalable and
cost effective purification strategy for the production of adenoviral vectors
from a range of suspension cell lines, from bench through to large scale, cGMP
clinical manufacture. The design of a ‘plug and play’ platform process and
integration of microscale chromatography techniques have allowed the rapid
deployment of an effective adenoviral purification process to meet
demand.
11:55 – 12:20
High-throughput process development technology for design
of cleaning-in-place (CIP) protocols for chromatography media
Dr Anna Grönberg, GE Healthcare¸UK
Cleaning-in-place (CIP) of chromatography media is
important for the integrity and safety of the final biopharmaceutical product.
Efficient and media compatible cleaning procedures also increase the column
lifetime and thereby contribute to cost effective processes. We
have developed a methodology where numerous cleaning agents and sequences of
cleaning steps can be evaluated in parallel using PreDictor™ plates, i.e.
96-well filter plates pre-filled with chromatography media. The PreDictor
plates were cycled repeatedly with feed and the cleaning efficiency of a large
number of different chemicals and sequences of cleaning steps were evaluated by
analyzing the residual amount of proteins on the beads after cleaning. The
throughput of the method was maximized by implementing the workflow on a
robotic system and by using high-throughput analysis. The
correlation between the scale-down screening format and traditional column
lifetime studies will be discussed. Process economy calculations comparing
different resins and cleaning regimes will also be presented.
12:20 – 12:45 Talk
to be confirmed
TBC, Tecan Trading AG, Switzerland
12:45 –13:40 Lunch and Poster Viewing
13:40
- 14:35 Question and Answer Session
Delegates
will be asked to submit questions to a panel of experts. Questions can be submitted before the event
or on the day
14:35 - 15:00
Use of
high throughput process development for the process optimisation for Fab
antibody fragment purification
Dr Dev
Baines, ProMetic BioSciences Ltd
Engineered antibody fragments are of increasing importance as
next-generation antibodies for wide ranging applications as biopharmaceuticals
and diagnostic tools. The presentation will describe high throughput micro
scale downstream process development for purification of Fab antibody
fragment using a 96 well plate format
tool (PuraPlate™) for process optimisation in conjunction with an adsorbent
developed for the capture of antibody fragments (Fabsorbent™ F1P HF). This process was used to optimise for equilibration,
sample loading and elution strategies.
High throughput experiments allowed for a wide range of buffers and
additives to be evaluated in relatively
short period of time. The pH of the
elution buffer was identified as the most was important parameter for the
recovery of the Fab fragments from E. coli lysate with high purity
15:00 –
15:30 Afternoon Tea/Coffee and Last Poster
Viewing
15:30– 15:55 Talk to be confirmed
Dr Sam Denby, Oxford Biomedica,
UK
15:55 – 16:20
Engineering High-Throughput Formulation Development
Dr Yitzchak Grant,
UCL, London
Lyophilization formulation has often
been referred to as a "dark art." Formulations are selected empirically
based on sketchy scientific knowledge and hit and miss approaches. The aim of
this investigation was to engineer a systematic process that identified optimum
formulations using minimal quantities of material in an automatable platform
which would lend itself well to high throughput screening methods.
16:20 – 16:45 Engineering
characterisation of miniaturised systems as a basis for rapid bioprocess design
and scale-up
Professor Gary Lye, Professor of Biochemical
Engineering, University College London
Advances in the miniaturisation of bioprocess unit
operations were initially driven by the need for small scale cell culture
devices. These are now being matched by novel downstream processing
technologies, designed to operate at complementary scales, along with the
automation necessary to facilitate parallel and high throughput
experimentation. To proceed as rapidly as possible through the stages of
bioprocess creation, scale-up and validation, however, requires that the data
obtained at each step are quantitative and predictive of larger scales of
operation. This presentation will summarise our fundamental understanding of
miniaturised bioprocess operations and how this can be used to obtain the
greatest benefits from investment in these technologies. Examples will cover
aspects of scale-up from cell culture through primary recovery to
chromatography highlighting some of the analytical and regulatory challenges
that remain to be addressed.
16:45 - 17:00
Chairman’s summing up
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About the chair
Gary Lye, FIChemE ( http://www.ucl.ac.uk/biochemeng/staff/lye.htm), is Professor of Biochemical
Engineering and Deputy Head of the Department of Biochemical Engineering at
University College London (UCL). He received his PhD in Biotechnology from the
University of Reading in 1992 and subsequently held posts in Chemical
Engineering at Imperial College London and the University of Edinburgh before
joining UCL in 1996. He has broad research interests on the application of
microscale and automation techniques to the rapid design, optimisation and
scale-up of bioprocesses. At UCL he is Director of the EPSRC Industrial
Doctoral Training Centre (IDTC) in Bioprocess Engineering Leadership and a
member of the Innovative Manufacturing Research Centre (IMRC) in Bioprocessing.
He was also a member of the recent UK Government Industrial Biotechnology
Innovation and Growth Team.
About the
speakers
Jonathan Dempsey is a Process Science Fellow for
Invitrogen’s PD-Direct Services, joining Invitrogen in 2007 after over 15 years
spent working in the Biopharmaceutical industry, where he gained tremendous
experience in the development of microbial and cell culture manufacturing
processes and cell lines. Jon has also been directly involved in the
development of several commercial biotherapeutics. Jon’s current
responsibilities are to advise and assist European biotechnology
companies in developing products and processes for the manufacture of
biotherapeutic manufacturing processes.
Ray Field is Director of Cell Sciences, in the
Development department at MedImmune (formerly Cambridge Antibody Technology)
the biologics arm of AstraZeneca. He currently leads development
functions including cell line development and upstream bioprocessing
including, in-process assays at MedImmune’s Cambridge R&D site.
He has previously held scientific and leadership positions at Celltech
Biologics and also in the research division of (Astra)Zeneca
Pharmaceuticals
Jonathan Souquet is working as a Senior Downstream
Process Development Scientist at Eden Biodesign developing and optimizing
purification strategies for a wide range of bio-molecules including recombinant
proteins, antibodies, viruses and virus like particles. Jonathan also plays a
key role in the subsequent scale up and tech transfer of processes into the GMP
processing facilities at the National Biomanufacturing Center and in providing
technical support during manufacturing activities. Prior to Eden Jonathan
attained a Ph.D. in the field of chromatography from the department of
Biochemical Engineering at the University of Birmingham working within the
bio-separations research group. He was also awarded a M.Sc. in
Biochemical Engineering from University College London.
Anna Grönberg who has a M.Sc. in Molecular Biotechnology has worked at
GE Healthcare Life Sciences R&D since 2003. She has been involved in the
development of state-of-the-art chromatography media such as MabSelect SuRe™
and Capto™ adhere by performing application oriented work. Lately, Anna has
focused on the challenge of developing efficient cleaning-in-place protocols.
She has developed a high-throughput work flow to screen and optimize CIP
protocols for specific chromatography steps in a process. This technology will
help producers of biopharmaceuticals to significantly optimize the utilization
of chromatography media and thus improve process economy.
Kenneth Lee
studied in Birmingham university during both his undergraduate and
postgraduate. Whilst at Birmingham, Kenneth gained experience in various
academic and industrial laboratories including Loughborough University, Keele
University, and Smith&Nephew, York. Kenneth
Lee started working at The Automation Partnership, a leader in the design and
production of automated cell culture since 1989, in 2009 as a Product
Development Engineer and Product Specialist in various projects, notably for a
new micro-scale bioreactor platform; more information on this system will be
given at the presentation. Kenneth has
experience in both process design, including industrial bioreactor design, and
practical cell biology and cell culture. His experience in both these areas
allows him to appreciate problems such as scalability of bioreactor system as
well as the practicalities of performing routine cell culture and cell
physiology.
Yitzchak Grant recently completed his EngD in biochemical
engineering at University College London. He spent four years developing a
systematic step by step screening and optimisation process for the development
of suitable biopharmaceutical formulations predominantly for lyophilization. He
also developed a method of empirical modelling of the lyophilization cycle
itself specific to the product being freeze dried.
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This meeting is
supported by
POSTERS
"EXPANSION
OF ENDOTHELIAL PROGENITOR CELLS (EPC) UNDER SERUMFREE CONDITIONS FOR CELL THERAPY IN PANSYS3000"
Wolfgang Erl (1), Josef Seidl (1), Michael Wiechmann (2)
(1) : PAN-Biotech GmbH, Germany
(2) : PAN-Systech GmbH, Germany
AUTOMATED EVALUATION OF MICROSCALE
LINKED PROCESS SEQUENCES FOR GENERATION OF SCALEABLE BIOPROCESS DESIGN DATA
J.Z. Baboo1,
J.M. Ward2, G.J. Lye1, M. Micheletti1
Department of Biochemical
Engineering1 & Institute of Structural and
Molecular Biology2, University
College London
Torrington Place, London WC1E 7JE
UK
Corresponding author: m.micheletti@ucl.ac.uk
Oxidative bioconversions offer valuable opportunities
in industrial pharmaceutical synthesis such as using Baeyer-Villiger
monooxygenases for antibiotic synthesis1. However, a limiting factor
is the identification of scaleable hydroxylation biocatalysts. Coupling
high-throughput microscale techniques with automation enables the operation of
linked process sequences for faster identification and characterisation of
optimal conditions2. A fully automated microscale sequence involving
fermentation, induction and bioconversion has been developed for the evaluation
of whole cell Baeyer-Villiger monooxygenases. The automated approach has been
shown to be robust and reproducible over multiple runs producing consistent
results on different days. Rapid automated collection of quantitative kinetic
data on new bioconversion substrates, substrate concentrations, media formulations
and well fill volumes has been achieved. By using a matched oxygen transfer
coefficient (kLa) approach both fermentation and bioconversion
operations have been successfully scaled up to 2 L3 and 75 L scale.
Current research is focusing on applying the automated microwell sequence for the
study of P450 enzyme-catalysed bioconversions4.
References:
1. Strukul G (1998) Transition Metal Catalysis in the
Baeyer-Villiger Oxidation of Ketones. Angewandte Chemie International Edition
37:1198-1209
2. Lye GJ, Ayazi-Shamlou
P, Baganz F, Dalby PA, Woodley JM (2003) Accelerated design of bioconversion
processes using automated microscale processing techniques. Trends in
Biotechnology 21:29-37
3. Ferreira-Torres C, Micheletti M, Lye G (2005)
Microscale process evaluation of recombinant biocatalyst libraries: application
to Baeyer–Villiger monooxygenase catalysed lactone synthesis. Bioprocess and
Biosystems Engineering 28:83-93
4. Hussain HA & Ward JM (2002) Enhanced
heterologous expression of two Streptomyces
griseolus cytochrome P450s and Streptomyces
coelicolor ferredoxin reductase as potentially efficient hydroxylation
catalysts. Applied and Environmental Microbiology 69: 373-382
SCALE DOWN
BIOPHYSICAL ANALYSIS FOR BIOPHARMACEUTICAL PRE-FORMULATION
Simon Webster
Avacta Limited, UK