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C1, An Ultra-High Yielding, Game Changing Gene Expression Platform
December 21, 2017
DYADIC INFORMATION
Safe Harbor Regarding Forward-Looking Statements
Certain statements contained in this presentation are forward-looking statements within the meaning of the federal securitieslaws. These forward-looking statements involve risks,uncertainties and other factors that could cause Dyadic’s actualresults, performance or achievements to be materially differentfrom any future results, performance or achievements expressedor implied by such forward-looking statements. Any forward-looking statements speak only as of the date of this presentationand, except as required by law, Dyadic expressly disclaims anyintent or obligation to update or revise any forward-lookingstatements to reflect actual results, any changes in expectationsor any change in events. Factors that could cause results to differmaterially are discussed in Dyadic’s publicly available filings,including information set forth under the caption “Risk Factors” inour December 31, 2016 Annual Report filed with OTC Markets onMarch 24, 2017. New risks and uncertainties arise from time totime, and it is impossible for us to predict these events or howthey may affect us.
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DYADIC INFORMATION
Table of Contents
Section Page
Title 1
Safe Harbor Regarding Forward-Looking Statements 2
Table of Contents 3
Dyadic Overview 4
C1 Production Host 8
C1 For Biologics 11
Glycoengineering 18
Advantages of using C1 for the Development & Production of Biologics and Vaccines 21
Summary 27
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DYADIC INFORMATION
Dyadic is Developing What the Industry Refers to As a “CHO stopper”
CHO stopper? Biogen looks to alternative cell lines for future of bioproduction
The Chinese hamster ovary (CHO) cell line is not the future for biomanufacturing says Biogen, MIT & Gates Foundation
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BioPharma Reporter Bioprocessing survey report, 11/03/2017
“Nearly half the respondents of our second state of the global biomanufacturing survey believe we are too reliant on Chinese Hamster Ovary (CHO) expression systems.”
to further develop C1 into a safe and efficient gene expression system to help speed up the development, lower production costs and improve the performance of biologic vaccines and drugs at flexible commercial scales.
Dyadic’sGoal
DYADIC INFORMATION
Platform TechnologyC1: Fungal Gene
Expression Platformfor use in the Development and
Production of Biologics
HQ: Jupiter, FL
BD&L: London &Budapest
R&D: Finland& Spain
1979 FOUNDED
Value & Differentiation:Decreased
Development TimeLower
Production CostsImproved
Biologic Performance
>20 Patents
Novel engineered cell line (Myceliopthora thermophila)
20+ YEARS EXPERIENCE IN PHARMA / FUNGAL GENE EXPRESSION PLATFORMS
GRAS FDA Certified
500,000LScale Production
>100 g/l Yield &~80% Purity
Hyper Productive Enzyme Expression
Industrial Licensees:DuPont, BASF, Abengoa, CODEXIS, Shell etc.
Dyadic Overview
Industrially Proven
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DYADIC INFORMATION
Dyadic Leadership Team & Financial Overview
Leadership Team Financials
$75MDeal with DuPont for Dyadic’s Industrial Technology Business
>$110MC1 Related License Deals, Milestones & Equity
$51M Cash & Investment Grade Securities (1)
$0Debt
M. Emalfarb Founder, CEO
$44M Market CapOTC Markets Stock Exchange(OTCQX: DYAI)
$19M Share Buyback Completed 2/2017
$5M Add’l Share Buyback Initiated 8/2017
R. Tchelet, PhDVice President, R&D
M. JonesCommercial Officer 28.7M
Common Shares Outstanding (1)
T. DubinskiVice President, CFO
6(1) As of September 30, 2017
Liquidity
Fully Funded to Execute Business Plan
DYADIC INFORMATION
Dyadic Board – Decades of Big Pharma Experience
Board of Directors Experience
Senior Vice President in Pfizer’s US Pharmaceutical Division.
Dr. Bose worked at Pfizer for 34 years and held leadership roles within bioprocess development and clinical manufacturing and is widely recognized as a Key Thought Leader in the biopharmaceutical industry.
Arindam Bose
Mr. Tarnok spent the majority of his career at Pfizer and is a seasoned finance and operational executive with extensive pharmaceutical industry experience.
Currently serves on the Board of the Global Health Council, and Ionetix, Inc. Prior Board service includes Keryx Biopharmaceuticals, Inc., where he also served as Chairman of the Board.
Michael P. TarnokChairman
Vice-President, Biotherapeutics Pharmaceutical Sciences, External Affairs and Biosimilar Strategy
Last Position
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DYADIC INFORMATION
C1 Production Host
Reinventing biological vaccine and drug development & productionDYADIC®
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DYADIC INFORMATION
CHO C1 White Strain 2.0
C1 – The Science
Unique Morphology
High Purity - 80% of target protein secreted
Wide operating conditions for pH and temperature
Shorter Development & Production Cycle
Translates into better growth conditions• Higher yields of secreted protein• Lower viscosity
Greater retention of target secreted protein through downstream processing
Requires only low cost synthetic media No Viruses which eliminates 2 purification
steps typical in CHO• No Low pH viral inactivation• No Virus nanofiltration
At scales ranging from laboratory shake flasks to 20,000l tanks and above
C1 has received GRAS (Generally Recognized as Safe) designation from FDA and is considered fit for human consumption
Develop g/l/d C1 cell lines in 15 weeks From seed flask to fermenter
• Savings of nearly 10 -14 days vs CHO Fermentation Cycle time 4-7 days
• 1/2 to 1/3rd the time of CHO
Temperature
40ºC37ºC
-32ºC25ºC
CHO
3
45ºC-
25ºC
C1
71 14
5 8CHOC1
pH1 7
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DYADIC INFORMATION
• High Protein Expression: • The C1 cell line achieved productivity up to 80 g/l of the target protein/enzyme
Fermentation Profile of Total Protein Production by HC strain vs. Single Proteins Production by LC strain
C1 Fungal Gene Expression Platform: World Class Expression
C1 Production strain = HC strain
C1 White strain = LC strain
LC strain expressing Indigenous enzyme for licensee partner
LC strain expressing heterologous enzyme by licensee partner
(1)
(2)
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DYADIC INFORMATION
C1 for Biologics
Reinventing biological vaccine and drug development & productionDYADIC®
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DYADIC INFORMATION
We have expressed 100% of the mAbs tested in our 3rd party research collaborations Initial unoptimized level of mAbs usually reach 2-5 g/L in 4-7 days fermentation Highest level of unoptimized expressed mAb is 1.22 g/l/d, (Fc-fusion – 1.35 g/l/d) The mAbs are integrated specifically to a “Hot spot” in the genome After the integration the selective marker is being eliminated The mAbs are secreted to the media and are being properly folded
High Yield & Purity of C1 Expressed mAb’s
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DYADIC INFORMATION
A certain Mab for which the ligand was commercially available was produced in CHO (control Mab) and C1 (C1-produed mAb)
The binding properties of the mAbs to the ligand were compared in a Biacore T200 assay
The control mAb and C1-produced mAb showed virtually indistinguishable binding kinetics.
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• The Binding Kinetics of C1 and CHO produced mAbs appear near identitcal
C1 mAb’s: Virtually Indistinguishable Binding Kinetics to CHO
DYADIC INFORMATION
Agglutination test
Influenza strain Expression Bioactive HA
New Caledonia, A (H1N1) Yes Yes
Texas, A (H1N1) Yes Yes
Puerto Rico A (H1N1) Yes Yes
California, A (H1N1) Yes Yes
Florida B Yes Yes
C1, Ability to Express Biologically active HA’s
The Expression of 5 Recombinant HA’s by C1
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DYADIC INFORMATION
830
453
104108
HA - C1
1 3.33 10 30
HI t
iter
agai
nst
Infl
uenz
a vi
rus 1000
100
10
530 30 0
C1Mock1
C1Mock1 PBS
Negative control
μg HA μg HA
HA-C1 Excellent Immunogenic Properties
The full length rHA from A/New Caledonia/20/99 (H1N1) strain showed excellent immunogenicity properties in mice without adjuvant
Excellent Immunogenicity from C1 Expressed HA
Mice study was conducted by Sanofi-Pasteur
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DYADIC INFORMATION
Additional C1 HA Data from Mice Study Conducted by Sanofi-Pasteur
The full length recombinant HA produced in C1 did not induce any negative clinical signs in mice.
No weight loss. No negative clinical signs during the experiment (visual observations taken each day).
The full length of HA/New Caledonia produced in C1 showed excellent immunogenic properties in mice.
C1 can potentially produce levels of 1 g/L of HAs and other antigens in 4 - 7 days fermentation therefore:
In seasonal Influenza Vaccine—total doses distributed = 146M/year Each 0.5 mL dose is formulated to contain: 15 µg of HA for each strain. Thus, 3 X 1000L scale fermentation runs will be able to supply the annual global
HA/strain needs against Influenza of 2,175 g.
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DYADIC INFORMATION
ZAPI Project
ZAPI, is a research and development program sponsored by the EU with the goal ofdeveloping a platform suitable for the rapid development and production of vaccines andprotocols to fast-track registration of developed products to combat epidemic Zoonoticdiseases that have the potential to effect the human population.
GOAL
Three of the initial antigens, each one for a different virus, wasexpressed by C1 and secreted to the medium
To date one of the C1 expressed antigens was tested in a very smallmice test within the ZAPI project. Preliminary results indicated thatthe C1 produced antigen generated an immune response in micethat protected the mice, and did not have negative effects on thehealth of the mice
We have initiated a C1 development program to express Virus likeparticles (VLP) for antigen expressions
Nano-particleExpression molecule
BRUNE KD et al., Bioconjug Chem. 2017
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DYADIC INFORMATION
Glycoengineering
Reinventing biological vaccine and drug development & productionDYADIC®
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DYADIC INFORMATION
Glycoengineering of C1 strain will provide the formation of various glycan structures to evaluate immunogenicity
C1 typical Glycan structure
C1 Glycoengineering
Unlike most fungi and yeasts, C1 does not have ‘high’mannose (branched 30-50 mannose species), but ratherhas ‘oligo’ mannose and hybrid-type structure.
The native C1 glycan pattern is relatively complex withhigh mannose type (Man3-Man9) and hybrid type(Man3HexNac-Man8HexNac) glycan forms
So far, O-glycosylation was not identified in therapeuticproteins expressed in C1 but minor level is still possible
C1 future Glycostructures
Glycoengineering work is being applied to C1strain to create a strain that produce proteinswith defined human glycoforms
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DYADIC INFORMATION
C1 Glycoengineering In Progress
Dyadic’s C1’s glycan structure is moremammalian like than typical yeast
o The native C1 glycan pattern isrelatively complex with highmannose type (Man3-Man9)
o O-glycosylation was not identified intherapeutic proteins expressed in C1
o Less engineering steps needed for C1
The first steps of Glycoengineering C1cells has begun and were successful
No negative effects on cell viability havebeen observed with any of themodifications done
Dyadic C1 Glycan Structure
Typical Yeast Glycan Structure
Man9
Targeted Mammalian Glycoforms
G0 G0F G2 G2F
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DYADIC INFORMATION
Advantages of Using C1 for the Development & Production of Biologics and Vaccines
Reinventing biological vaccine and drug development & productionDYADIC®
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DYADIC INFORMATION
Better:• High Productivity Protein Expression: ~80 g/l• High Purity Protein Secretion (~80%)• Low viscosity• Greater Retention of target secreted protein through downstream
processing• C1 current developed strain can be used as production platform for
non-glycosylated proteins such as Fabs, bi-specifics and new drugs• C1 secreted therapeutic proteins do not require deletion of the O-
glycosylation machinery• Glycoengineering work is being applied to the host production C1 strain to
allow for production of proteins with human defined glycoforms such as mAbs, Fc-fusions and recombinant vaccines
Easier:• Advanced Genetic Tool Box• Site specific integration vs. random integration• Wide operating conditions for pH and temperature• Simple C1 production process allows for production of biologics at various
scales and at different sites
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C1 Advantages for Developing & Producing Biologics
DYADIC INFORMATION
C1 Advantages for Developing & Producing Biologics
Faster:• Develop high yield g/l/d C1 cell lines in 15 weeks
• Fed batch technology – no need for perfusion
• No Viruses eliminates the need for two additional purification steps
• 4-7 days Fermentation time (1/2 to 1/3rd less time than CHO)
• Cell Reproduction rate (2x greater than CHO)
• Initial protein production rate is (~1.5x greater than CHO) and expected to increase further
Lower Production Cost:• Defined, low-cost media based
on glucose
• No Viruses eliminates associated costs
Comparative Manufacturing Cost: C1 vs CHOHumira mAb
CHO - 10,000L tank (3)
Cost
in M
illio
n U
SD
21
*
C1 - 2,000L tank(1)
C1 - 10,000L tank (2)
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DYADIC INFORMATION
C1 Advantages for Vaccine Development & Production
Productivity – C1 is a highly productive strain that can produce rVaccines at very low cost.
• Example: C1 can potentially produce levels of 1 g/L of HAs against seasonable Influenza virus(es) in 4 - 7 days fermentation therefore:
In seasonal Influenza Vaccine—total doses distributed = 146M/year Each 0.5 mL dose is formulated to contain: 15 µg of HA for each strain. Thus, 3 X 1000L scale fermentation runs will be able to supply the annual
global HA/strain needs against Influenza of 2,175 g.
Flexibility– The relative simplicity of the production process of C1 enables the production of rVaccines at various scales and at different sites.
Safety – Mice tests demonstrated that recombinant proteins such as HA produced in C1 did not induce any negative clinical signs in mice.
No weight loss. No negative clinical signs during the experiment (visual observations taken each day).
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DYADIC INFORMATION
Immunogenicity – Antigens produced by C1 demonstrated excellent immunogenicity properties:
• Sanofi Project: The full length rHA from A/New Caledonia/20/99 (H1N1) strain showed excellent immunogenicity properties in mice without adjuvant
• ZAPI Project: C1 produced antigen generated an immune response in mice that protected the mice and did not have negative effects on the health of the mice
Adjuvant effect – Reducing rVaccines risk. Samples prepared by C1 possess Adjuvant properties. Thus, antigen produced by C1 may not require the addition of artificial Adjuvants
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C1 Advantages for Vaccine Development & Production
DYADIC INFORMATION
Proteolytic Activity
High Low
Proteolytic Activity
High Low
0 g/L 20 g/L
Basic Therapeutic Protein Productivity
0 g/L 20 g/L
Basic Therapeutic Protein Productivity
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• Current Production Strain
• Target C1 Production Strain
• Continuing to Optimize Yield, Stability, Glycan Structure & Purity Changing the
cellular regulatory circuit
Libraries of Efficient & Strong
Promoters
Libraries of TF and signal peptides and / or carrier proteins
Libraries of protease deletion
strains
Glycoengineering to form mammalian-like
glycan structures
As Good As C1 Currently Is, We’re Making C1 Better Every Day!
Protein Purity
DYADIC INFORMATION
C1 Benefits: Lower Production Costs, Both CAPEX and OPEX
2,000 liter2 x12,000 liter
C1 can lower CAPEX:• Produce at smaller
scale while dramatically increasing protein yields
C1 can lower OPEX• Smaller facility
footprint and related costs
• Low cost media
Single Use BioreactorStainless Steel Multiuse
CHO C1Annual Protein Demand in g 800,000 800,000 800,000
Tank size in Liters 12,000 2,000 2,000 Productivity g/l 3 10 15 % Yield 65% 75% 75%Batches per year 20 40 40 Tank Output in g 468,000 600,000 900,000 Tanks Needed 2.0 2.0 1.0 % Capacity Utilized 85% 67% 89%
DYADIC INFORMATION
Dyadic is looking for partners in the biopharmaceutical space to exploit the potential of C1. Contact mjones@dyadic.com
Ongoing Internal & Third Party Research Programs:
Optimizing Yield, Stability, Glycan Structure, Other Properties & Purity
Higherprotein yields
Lower CapEx/OpEx
Higher purity & greater protein
recovered
Low Cost Media / No
Viral Inactivation
No negative clinical signs
in mice studies
Shorter development & production
cycles
Summary
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R&D Collaborations Licensing Arrangements
Other Commercial Opportunities
C1 Advantages
THANK YOU!
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