providing single use bioreactors for the modern age
TRANSCRIPT
Providing single use bioreactors for the modern age
Business Model
Cellexus makes a range of disposable bioreactors
for the production of recombinant proteins from bacteria, yeasts and Mammalian cells.
BiopharmaceuticalsFood additives
CosmeticsBiofuels
Worldwide sales dynamics
Increasing trend towards use of disposables
Cost savingsReduced downtime
Reduced risk of contamination
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.
Worldwide sales
2013 2014 2015 2016 20170
100
200
300
400
500
600
700
Worldwide market for GMP Disposable Biore-actors ($m)
WW Marketsize
Worldwide sales (fermentation)
21032014
20152016
2017
0
50
100
150
200
250
Disposable fermentation reactor market worldwide $m
CellMaker
Touchscreen User interface
Communications + sensor readers
Gas supply + Mixing + temp control
RegularPlusFelix
8L & 50L sizes
Sterile single-use bags
How it works
Competition: Microbial
These customer driven USPs are evident upon comparison against competitor products in the field of microbial cell culture:
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
Technical Information
CellMaker systems – flexible layout
Step 1 – Menu screen
Step 2 – change parameters
CellMaker PLUS Controller: Software
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CellMaker Applications
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
• 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
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)
Shaker flask, Regular and Plus
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)
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
• 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
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
June14
Thank you for your attention
Cellexus Ltd, Albacom, George Buckman Drive, Dundee, DD2 [email protected]