providing single use bioreactors for the modern age
TRANSCRIPT
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Providing single use bioreactors for the modern age
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Business Model
Cellexus makes a range of disposable bioreactors
for the production of recombinant proteins from bacteria, yeasts and Mammalian cells.
BiopharmaceuticalsFood additives
CosmeticsBiofuels
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Worldwide sales dynamics
Increasing trend towards use of disposables
Cost savingsReduced downtime
Reduced risk of contamination
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Cost comparisons
stainless steel
single use
-30%Electricity -38%
Space
-62%Steelwork
-87% Water
-95%CleaningMaterial
Reference: Sinclair A, et al 2008. The environmental impact of disposable technologies. BioPharm International Supplements.
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Worldwide sales
2013 2014 2015 2016 20170
100
200
300
400
500
600
700
Worldwide market for GMP Disposable Biore-actors ($m)
WW Marketsize
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Worldwide sales (fermentation)
21032014
20152016
2017
0
50
100
150
200
250
Disposable fermentation reactor market worldwide $m
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CellMaker
Touchscreen User interface
Communications + sensor readers
Gas supply + Mixing + temp control
RegularPlusFelix
8L & 50L sizes
Sterile single-use bags
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How it works
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Competition: Microbial
These customer driven USPs are evident upon comparison against competitor products in the field of microbial cell culture:
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Ease of Use
• Sterile bag prevents cross contamination
• Simple set up and simple to use
• High OTR
• Accurate temperature control using Peltier plates
• Increased performance using enriched air
• Reduced and easily defined costs
• Small footprint
• Fast changeover
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Technical Information
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CellMaker systems – flexible layout
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Step 1 – Menu screen
Step 2 – change parameters
CellMaker PLUS Controller: Software
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14
CellMaker Applications
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CellMaker REGULAR Applications
• E.coli– Primary host cell for recombinant protein expression
• Scale-up of culture from shaker flask• Increased aeration
– Potential for higher ODs, and higher protein yields– Potential higher yield of poorly soluble protein
• Low temperature induction of protein expression– Potential higher yield of poorly soluble proteins– Potential higher yield of toxic proteins
• Pichia pastoris– Widely used methylotrophic yeast for protein production
• Above benefits (for E.coli) hold true for P.pastoris
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• Algae– Diverse group of microorganisms rich in nutrients,
proteins and active compounds• Scale-up of algae culture to support these biodiversity
profiling activities.• Development of recombinant protein expression
technologies in algae• Ability to grow phototrophically or heterotrophically
• Insect cell culture– Popular recombinant protein expression system
• Baculoviral infection of insect cells (Sf9, Sf21, Hi-5)• Suspension culture• Higher oxygen demand following baculoviral infection
CellMaker REGULAR Applications
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E.coli: Scale-up from shaker flasks in the CellMaker
0 45 105 135 180 240 300 3300
0.5
1
1.5
2
2.5
3
OD CellMakerOD Shaker
1mM IPTG
Time (min) OD CellMaker OD Shaker0 0.11 0.09
45 0.22 0.14105 0.61 0.54135 1 0.9180 1.7 1.6240 2.4 2.1300 2.7 2.3330 2.8 2.4 1 2 3
Target Protein
1: Uninduced2: CellMaker3: Shaker
3.5hr induction of protein expression in E.coli at 37°C
Data provided by S Whyte (TwistDX)
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Shaker flask, Regular and Plus
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P. pastoris : Improved protein yield in the CellMaker
• Enhanced aeration of media through sparge tube
• Oxygen enriched gas supply, maintained at 4L/min Oxygen + 2L/min Air
• Higher ODs obtained compared to shaker flasks
• 1.5-fold increase in biomass
• 2-fold increase protein yield
• Protein appropriately glycosylated and secreted
0 24 48 72 960
10
20
30
40
50
60
Pichia pastoris growth at 29°C
CellMaker RegularShaker
Time (hours)
OD
TIME OD (CellMaker) OD (Shaker)
0 0.94 0.94
24 40 33
48 49 33.8
72 50 32.5
96 54 33
Data provided by M Rossmann (LMB, Cambridge)
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S. Aureus Bacteriophage: Scale-up in the CellMaker
0 50 100 150 200 250 3000
0.1
0.2
0.3
0.4
S.aureus OD600
OD 600 (3)
Time (mins)
OD
600
Phage addition
Data kindly provided by Y Jia (Novolytics, University of Warwick)
TIME (mins)
OD600 Comments
30 0.09770 0.16290 0.26995 - 1% phage (80ml) added100 0.350 OD taken straight after
phage addition185 0.358 Sign of cell lysis255 0.046 Culture cleared; harvest
• Seed the CellexusBag with S.aureus
• Allow bacteria to progress to early exponential growth phase
• Inoculate with bacteriophage
• Phage-enriched media later harvested following successful phage infection and lysis
• Convenient, user-friendly, consistent system for scale-up of bacteriophage production
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• Growing algae utilises windows in CellMaker Enclosure
• Perspex panels allows light through but insulates CellexusBag
• Peltier heat pumps cool CellexuxBag to 26oC (or as low as 15oC)
Algae: Growth in the CellMaker
3
In collaboration with the Welding Institute, Abingdon, Cambridge
CellexusBagTM - the first Cellexus technology
2
3
1
Key to the effective
mixing by gas (airlift)
and propellers (STR) is
the geometry of the
bioreactor.
2 patent applications filed World-wide
Reflector
Low heat, high efficiency lightsource
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CellMaker System Features & Benefits
Features BenefitsSimple 30 minute set-up Saves time, simplifies trainingUnique airlift bags Gentle but effective aeration to improve
yieldsOxygen generator (up to 4L/min) Simple installation
Saves moneyNo oxygen bottles to handle
Peltier-driven cooling & heating Greater application scope (e.g. expression of toxic proteins in E.coli clones) Algae at 26oCInsect cell growth (e.g. at 27oC)
Accurate temperature profile (+/-0.1oC) Reliable processes which give optimal protein production
Efficient condenser Removes vapour and odour from exhaust gases, creates better user environment
Highly robust build quality Maximises uptime 8L & 50L units available Choice of enclosure sizes to suit batch Scalable design Flexible designSimple operator software Fast set up time, simplifies training
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June14
Thank you for your attention
Cellexus Ltd, Albacom, George Buckman Drive, Dundee, DD2 [email protected]
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