Integrated synbio tools to enable biopharma production systems · 2017-08-21 · enable biopharma...
Transcript of Integrated synbio tools to enable biopharma production systems · 2017-08-21 · enable biopharma...
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CONFIDENTIAL
Integrated synbio tools to
enable biopharma production
systems
Ian Fotheringham
Ingenza Ltd.
www.ingenza.com26th July 2017
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Ingenza Ltd
Capabilities: 41 staff: (chemists and bioscientists)
Strain construction / protein expression / enzyme evolution
Microbial fermentation / Cell culture
Bioprocess development, DSP, chemical synthesis
All integrated for Industrial Biotechnology / Synthetic Biology application
Customers/Partners where synthetic biology applies:
Chemical companies developing biobased chemical syntheses
Pharma companies outsourcing pathway optimization, enzyme evolution
Academic groups transitioning early stage research to POC and spinout
COGS, new product/functionality, IP protection, sustainability
New synthetic routes, COGS, product diversification…funding (SME)
PoC, feasibility, viability, funding (spinout)
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• For each target chemical, polymer intermediate or pharmaceutical
• Expression of target enzymes in novel pathways
• Rapid iteration to improve productivity
• Enzyme adaptation
• Confirmed process Feasibility, Economics and Scalability
Engineered pathways to high value chemical building blocks
v vvvB P
vv
Combinatorial assembly
‘Omics interpretation, Revision
PhylogeneticEnzyme Sampling
Screening / Process Testing
ScalableBioprocess
Pilot and Full ScaleManufacturing
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Delivering predictability through methodologyAdaptable hierarchical screening
Calibrated screen dev
eloped for biotransfo
rmation
Adapted to optimis
e
biologic synthesis
Progression to drug
substance manufactu
re
46,000 novel
codon revisions
ReporterLibraryHighest producers
identified
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inABLE® – Combinatorial gene assembly
Key advantages:
One-step assembly
No PCR (errors)
Re-usable library of parts
No purification, fully automated
Synergistic with versatile HTP screening (solid/liquid phase)
Part Cv Part DPart A Part B v
vPart Cv Part DBPart A P Part B
Part DvPart CvPart BPart A
Enzyme activity
Toxinresistance
Crossfeeding
Colorimetric
AutomatedLCMS
Design/preparation
Part/linker fusion
Pathway assembly
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POC for bioprocess patents to key intermediates
Target enzymes expressed
Biotransformations and products characterised
> 40 independent steps in novel configurations
Reactions exemplified … IP established
Enabled multiple development programs to targets
Feasibility
Viability
Scalability
Enzymatic activity determination/measurement
Enabling innovative bioprocess developmentBiobased chemical manufacture
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Delivering predictability through methodology‘Omics informed design and revision
X X
X X
Phylogenetics, metabolomi
cs
structural biology
Precise genome integr
ation
Reduced processing
Bioethanol efficiency s
avings
Hierarchical screen/t
est
v vvvB P
vvinABLE® assembly
Serine integrase
adapted to yeast
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Delivering predictability through methodologyPredicting optimal translation
Modeling heterolo
gous mRNA transla
tion
Assembly and testin
g with target biolo
gic
v v
vvB P
vv
kDa16011060
40
3020
1510
20% uplift
> 9 g/L
5 L fed-batch fermentor validation
High volume/high value product –now comme
rcial
Aberdeen University, UK.Prof. Ian StansfieldProf. Carmen Romano
“tRNA abundance/ribosome hopping”
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Target protein almost completely insoluble in E.coli:
Resistant to all attempts to achieve soluble production
Unaffected by host tRNA adaptation, shake flask or fermentation conditions
No activity in cell extracts
Target
Insolublefraction
Solublefraction
Efficient protein expression systemsHorses for courses
Optimised Pichia pastoris production system:
No 3rd party IP, cGMP compliant
Efficient, high cell density fed-batch
fermentation
Product secreted, soluble and active, now
commercial
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Ingenza and Plymouth University researchers
• nM potency for Gram + pathogens (MRSA)
• Nasal decolonisation market
• Single dose efficacy vs (15 dose ) market leader
cfu/n
ostri
l
U n tre a te d
V e h ic le
N 1 0 1 , 0.8 % , O
N C E
N 1 0 1 , 0.0 4 % , B
ID
N 1 0 1 , 0.2 % B ID
Mu p iro c in , 2
% B ID1 0 0
1 0 1
1 0 2
1 0 3
1 0 4
1 0 5
1 0 6
1 0 7
L o g R = 2 .1 0 .0 9 0 .0 6 2 .5 9
Untreated Control NI01
Single dose
Antimicrobial discovery / production / engineeringEpidermicin NI01 bacteriocin
Highly promising product class
Low native yield
Adaptable target range:
Potency towards Gram - pathogens
J Antimicrob Chemother (2017) 72 (3): 778-781.
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> 50x yield
1 2 3 4 5
6 M
Fully active
$1.5 M funding for process scale-up and peptide adaptation
• Spinout (“Kaiju”) progressing lead compound to toxicology
• Machine learning informed adaptation to enhance epidermicin properties: Ingenza-Plymouth-NPL-STFC-IBM
• Solid/liquid phase screens in P. pastoris
Gene configuration, integration, induction
Antimicrobial discovery / production / engineeringEpidermicin NI01 bacteriocin
> 50x productivity, viable, scalable, fermentation protocol
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Need and opportunity :
500,000 (WHO) untreated haemophiliacs (75%) mainly in poorer nations
Requirement for low cost recombinant Factor VIII
Current cell line yield unable to support low cost market entry
$8 BN market today
Partnership of ProFactor Pharma and Ingenza: synergy of know-how / assets
High producing Factor VIII cell l ine, process
Identify novel construct and manufacturing process
Factor VIII
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Codon optimised FVIII - CHO
Proprietary FVIII +VWF chaperone
Efficient, scalable SUF perfusion protocol
Proprietary DSP
10x industry standard productivity FVIII
+
B-domain
deletion
High producing Factor VIII cell l ine, process
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Ingenza:
Worldwide partnerships with leading manufacturers
Enabling new products /markets /functionalities / University spinouts
Adaptable enabling technologies:
Combinatorial inABLE® genetic assembly
Gene design, targeted gene integration, ‘Omics
Scalable fermentation bioreactor protocols including single-use
Our Technology and Business focus is on:
Long-term high-value relationships
Scalable, cost-effective, sustainable bio-manufacturing
Synthetic Biology design plus proven Industrial Biotechnology methods
Bioprocess efficiency Requires “Abilities”(feasibility, adaptability, predictability, scalability)