Dyadic International, Inc. · NASDAQ:DYAI
Dyadic International — AVMI Webinar (June 2021)
47 slides from Dyadic International's conference presentation, June 2021
Slide 1: Making Healthcare Accessible & Affordable to Patients Globally
MAKING HEALTHCARE ACCESSIBLE & AFFORDABLE TO PATIENTS GLOBALLY Dyadic Thermophilic Filamentous Fungus (C1-cells) Recombinant Glycoprotein Antigen Vaccines, Antibodies & Other Therapeutic Protein Product Production Platform
Slide 2: Safe Harbor Regarding Forward-looking Statements
Safe Harbor Regarding Forward-looking Statements Certain statements contained in this presentation are forward-looking statements within the meaning of Section 27A of the Securities Act of 1933 and Section 21E of the Securities Exchange Act of 1934, including those regarding Dyadic’s expectations, i
Slide 3: Company highlights
Company highlights Next Generation Protein Expression Biotech with Well-established Global Partners Proprietary & Patented C1 gene expression Designed to bring biologic vaccines, drugs and other biologic products to platform technology market faster, in greater quantities, at lower cost, using micro
Slide 4: C1 gene Expression Platform
C1 gene Expression Platform 4
Slide 5: Filamentous Fungus C1-Cell High Level Protein Production
Filamentous Fungus C1-Cell High Level Protein Production High Purities & Exceptional Yields/Stabilities with Scalable Benefits for Protein Based Product Manufacturing C1-cells are thermophilic filamentous fungi with highly desirable growth and manufacturing properties C1 initially developed to produ
Slide 6: C1 is a safe production cell line
C1 is a safe production cell line Dyadic's GRAS Notice – C1 was acknowledged as a safe microorganism for the production of A Dyadic cellulase enzyme for food and feed application by the FDA on September 29, 2009. Non-Pathogenic Ø Strain is non-toxic and non-infectious Ø No toxic byproducts are gener
Slide 7: C1 Site Directed Transformation Method
Company highlights C1 Site Directed Transformation Method leads to quick generation of stable C1 cell lines Rapid Response Original gene The same rapid New gene High Doses method will be • Set of strong promoters Scalability needed to native and synthetic Affordability construct new • No need for in
Slide 8: C1 Fermentation Technology
C1 Fermentation Technology • Fully defined synthetic medium Fed-batch Process • Fed-batch technology with glucose feeding • Wide range of conditions available pH: 5-8, Temp: 48 hrs 24 hrs 120-168 hrs 20 - 45°C Seed flask Fed-Batch NH4OH for pH control • Low viscosity culture due to unique morphology
Slide 9: Certolizumab production with C1 in Single use Bioreactor
Certolizumab production with C1 in Single use Bioreactor (SUB) Conditions A GE’s Xcellerex™ XDR-50 MO Conditions B 12 . 12 SS-B SS-A 10 SU-B 10 SU-A 8 Fab (g/L) 8 Fab (g/L) 6 6 4 4 2 2 0 0 0 20 40 60 80 100 120 0 20 40 60 80 100 120 Age (h) Age (h) Comparison of Fab production kinetics in either SSB
Slide 10: Generic Process Flow Chart for C1
Generic Process Flow Chart for C1 (12 – 14 days) (*) Pre-inoculum: Passage 1 1 – 1,5 days Mycelium activation plate Passage 2 Seed train: inoculum flask expansion in flak 0,75 -2,5 days N-3 ~1.6L scale Inoculum expansion - bioreactors N-2 ~40L scale N-1 ~1000L scale 5 days (*) Generic Process Flow C
Slide 11: Summary - Titers Achievable in the C1 Host
Summary - Titers Achievable in the C1 Host Trispecific Fc-Fusion mAbs Fab (Certolizumab) up to 6.12 g/l 144 Hours 15.3 g/l up to 24.5 g/l up to 14.5 g/l up to 1.02 g/l/day 168 Hours 168 Hours 164 Hours Bispecific 2.58 g/l/day up to 3.5 g/l/day up to 2.1 g/l/day up to 6.17 g/l 72 Hours up to 2.05 g/l
Slide 12: C1-Cell Recombinant Protein Products With Competitive Advantages
C1-Cell Recombinant Protein Products With Competitive Advantages Robust Recombinant Protein Production Platform Offers Competitive Advantages Over Existing Techs High retention of target secreted protein(s) or other C1-cell bioproduct(s) through Purities downstream processing No viral or endotoxins
Slide 13: Vaccines
Vaccines 13
Slide 14: C1 is ideally suited to help meet global demand for Health Equity
C1 is ideally suited to help meet the global demand for Health Equity including multivalent vaccines & multi antibody cocktails Building a more efficient biomanufacturing platform to serve a global population more affordably Opportunistic participation in product development of vaccines, biotherapeu
Slide 15: EU Zoonosis Anticipation Preparedness Initiative ZAPI Final Conference
European Union Zoonosis Anticipation Preparedness Initiative ZAPI Stakeholders Final Conference Dyadic’s C1 Gene Expression Platform Played a Key Role In The Success Of The Five-Year EU € 20 Million ZAPI Program • Many infectious diseases, including influenza and "Within the collaboration, we helped
Slide 16: ZAPI Stakeholders Final Conference. Evidence for C1 Approach
European Union Zoonosis Anticipation Preparedness Initiative ZAPI Stakeholders Final Conference. Evidence for C1 Approach Dyadic’s C1 Gene Expression Platform Played a Key Role In The Success Of The Five-Year EU € 20 Million ZAPI Program "Within the collaboration, we helped to validate a prolific fu
Slide 17: C1-cells selected for antigen manufacturing
ZAPI Stakeholders Final Conference C1-cells selected for antigen manufacturing “Efficient manufacturing processes are being developed, I Specifically would like to mention Dyadic, the C1 the fungal process there which gives enormous, enormous amounts of antigens and I think especially in the face of
Slide 18: ZAPI Stakeholders Final Conference
ZAPI Stakeholders Final Conference "However, If you look at the subunit production that was just also described by Ronen, It is pretty clear that the C1 expression, or using the C1 platform is the key advantage here for as is described for yields but also DSP purification, we don't have to compare t
Slide 19: Success in Expressing High Level of SBV & RVFV Antigens
Success in Expressing High Level of SBV & RVFV Antigens ZAPI, is a research and development program sponsored by the EU with the goal of developing a platform suitable for the rapid development and production of vaccines and protocols to fast-track registration of developed products to combat epidem
Slide 20: The RBD: 'Achilles heel' of the spike protein
The RBD: ‘Achilles heel’ of the spike protein ACE2 receptor Analyses of the antibody repertoire of RBD-targeting infected patients antibodies are so show that the vast potent, as their majority of binding prevents (potent) virus interaction neutralizing with the host cell, antibodies target thereby
Slide 21: RBD has key advantages as a vaccine antigen
RBD has key advantages as a vaccine antigen Receptor binding domain (RBD) Advantages of minimization of spike protein Antigen antigen to the RBD: minimization • Exposure of key neutralizing epitopes on the RBD to the immune system that are hidden in the closed spike conformation • Efficient inductio
Slide 22: Successful Execution of Government Sponsored Collaborations
Successful Execution of Government Sponsored Collaborations Positions Dyadic to Enter Clinical Trials Israel Institute for Biological Research (IIBR) • Entered into initial collaboration January 2018 • Focused on advancing C1 expression platform for the development and manufacture of recombinant vac
Slide 23: SARS-CoV-2 Spike RBD Is A Key Target For Potent Neutralizing mAbs
SARS-CoV-2 Spike RBD Is A Key Target For Potent Neutralizing mAbs ➢ In ~2 months, we developed a C1 cell line expressing the Receptor Binding Domain (23kDa) of SARS-CoV-2 spike protein Receptor binding domain: ➢ C1 stable cell line was developed that expressed the RBD originally at a level of ~ 1 g/
Slide 24: C1 RBD Binds to all Epitopes
C1 RBD Binds to all Epitopes DYADIC INFORMATION 24
Slide 25: Evaluation Of RBD Produced By C1
Evaluation Of RBD Produced By C1 Biomolecular Binding Kinetics Assays: The equilibrium 01 01 dissociation constant (KD) of C1 SARS-CoV-2-RBD-Ctag Biosensor binding to recombinant hACE2 was calculated to be 4.9 Nm, which is comparable to that of the CHO SARS-CoV-2- RBD: 5.11 Nm. hACE2 receptor RBD An
Slide 26: Mice study with C1 expressed SARS-CoV-2 RBD vaccine candidate - Results
Mice study with C1 expressed SARS-CoV-2 RBD vaccine candidate - Results Mice study demonstrated that the C1-RBD induced neutralizing antibodies at high level. ELISA PRNT 128,000 1st 2nd 3rd 6 1st 2nd 2nd 3rd 6 32,000 22,627 log10 serum dilution 5 0 19 21 28 42 43 50 11,310 4 10,196 log10 NT50 5 3 8,
Slide 27: Challenge Test with Transgenic Mice study
Challenge Test with Transgenic Mice study with C1 expressed SARS- CoV-2 RBD vaccine candidate Challenge Mice study demonstrated that the C1-S-RBD induced full protection A. Vaccination of K18-hACE2 transgenic mice: B. 2 groups of transgenic mice were vaccinated with 20 µg of RBD-C formulated with Al
Slide 28: N-glycosylation pattern. NP-HPLC chromatography analysis
N-glycosylation pattern. NP-HPLC chromatography analysis 121.68 M10 M11 117.23 M12 125.79 RBD-His 112.19 106.52 M13 120.68 129.54 Saccharomyces M14 110.84 100.29 132.79 * not identified structures M10-M14: N-glycans bearing 10-14 mannose residues 102.07 106.17 99.12 RBD-His 97.45 96.78 104.45 100.28
Slide 29: N-glycosylation pattern. NP-HPLC chromatography analysis
N-glycosylation pattern. NP-HPLC chromatography analysis The protein sequence was verified and the correct formation of the four disulfide bonds was confirmed. Additionally, the molecular weight of the intact protein was confirmed since the theoretical and experimental values, considering the N-glyc
Slide 30: Analysis of released N-glycans by UPLC-FLR-QTOF-MS
Analysis of released N-glycans by UPLC-FLR-QTOF-MS Results for RBD-C-tag Component Observed Amount Mass error name RT (min) ( %) (ppm) M3* 10,58 0,91 1,3 M3 11,15 0,04 1 M3-HexNAc 12,06 1,09 -0,1 M4 13,95 27,47 1,6 M4-HexNAc 15,23 35,52 1,6 M5* 16,39 0,21 0,4 M5 16,95 0,3 0,8 M5_HexNAc 18,21 15,84 1
Slide 31: Phase I with C1-cell SARS-CoV-2 RBD Recombinant Vaccine in 2021
Phase I with C1-cell SARS-CoV-2 RBD Recombinant Vaccine in 2021 cGMP manufacturing, Fill/Finish, Toxicology, initiating and completion of an anticipated Human Phase 1 Clinical Trial Of a C1 produced SARS-CoV-2 vaccine candidate. Prove Safety & Efficacy In Humans Developed the C1-RBD Coordination of
Slide 32: DYAI-100 / SARS-CoV-2 RBD Vaccine Candidate (cGMP, Phase I)
DYAI-100 / SARS-CoV-2 RBD Vaccine Candidate (cGMP, Phase I) ➢ Meeting with Paul Ehrlich Institute (PIE) was held on April 26th and Final Minutes were received on June 2nd. ➢ Toxicology study o All animals have been vaccinated 4 times and were sacrificed. The dissections will continue till Monday – J
Slide 33: Commercial Scale Production of C1-RBD With and Without MPSP
Commercial Scale Production of C1-RBD With and Without MPSP Predicted C1-RBD fermentation capacities for different dose requirements based on 5 days fermentation at various scales Doses (30µg and 30µg) Doses (15µg+15µg) C1 productivity (2.0 g/L) 10M 100M 1000M 10M 100M 1000M Total volume (g) 600 6 0
Slide 34: C1-Cell Expressing antigens and neutralizing mAbs for SARS-CoV-2 vaccines
C1-Cell Expressing antigens and neutralizing mAbs for SARS-CoV-2 vaccines • SARS-CoV-2 proteins and neutralizing mAb successfully expressed by C1 • S-RBD • Wuhan, South African, UK & Brazilian C1 / RBD Ctag C1 / RBD-SpyTag Neutralizing mAb no CTag • S-RBD / Spy Tag (Nanoparticle) • Full Spike / Spy
Slide 35: Stable C1-RBD-SpyTag Production Without C-Tag
Stable C1-RBD-SpyTag Production Without C-Tag 1L scale fermentation • Developed stable C1 cell lines expressing RBD-SpyTag without C-Tag. 1 - Mw marker 2 – EFT 55h 8µl • Production level in 1L fermentor reached 0.7-0.8 g/L in 55hrs. 3 – F.T. (15ml) 20µl • Efficient and simple purification steps with
Slide 36: Production of SARS-CoV-2 vaccine Protein Variants
Production of SARS-CoV-2 vaccine Protein Variants VTT is expressing the following proteins in C1: 1. Variants of Concern • C1 strains have been engineered to express high levels of Receptor binding domains (RBDs) of the UK, BR and SA variants • Delta (Indian) Variant(s) are in progress • Initial fer
Slide 37: Alternative Adjuvant to Initiate Cells Response
Alternative Adjuvant to Initiate Cells Response Do we need alternative Adjuvants? AS04 Beginning in 2009, monophosphoryl lipid A (MPL) was used in one U.S. vaccine (Cervarix®); however, the vaccine is no longer available in the United States due to low market demand. This immune-boosting substance w
Slide 38: Establishing Global Presence with leading organizations
Establishing Global Presence with leading organizations Co-developing C1 enabled COVID-19 (plus variant) vaccines and/or boosters (i.e., tetravalent or quadrivalent COVID-19 vaccines) Dyadic and Medytox To Develop Vaccines Dyadic announces development Against COVID-19 Variants of COVID-19 Vaccine in
Slide 39: Establishing Global Presence with leading organizations
Establishing Global Presence with leading organizations ➢ Purified protein and/or supernatants and/or production strains of the C1 expressed SARS-CoV-2 RBD & Full Spike proteins have been sent in more than 30 shipments to more than 20 different collaborators ➢ New strains have been and are continuin
Slide 40: Establishing Additional Global Presence with leading organizations
Establishing Additional Global Presence with leading organizations Asia North America India South America Africa Australia (including Middle East) DYADIC INFORMATION 40
Slide 41: Expression of Haemagglutinin (HA) in C1
Expression of Haemagglutinin (HA) in C1 SS Signal Sequence (HA SS for L talentolae expression, C1 SS for fungi expression) Native HA (trimer) SS Haemagglutinin TM CD Transmembrane Domain Secreted HA (monomer) SS Haemagglutinin Tag Cytoplasmic Domain Tag 6XHistidine Secreted HA + sequence-induced SS
Slide 42: MAb overview
MAb overview 42
Slide 43: Biomanufacturing Capacity: New blockbuster mAbs could leave shortage
Biomanufacturing Capacity: New blockbuster mAbs could leave shortage C1-Cells Disrupting Monoclonal Antibody Development & Manufacturing • Three to four batches of C1 mAbs in same time as one batch of CHO mAbs • Produce greater quantities from each batch more affordably • Same binding and neutralizi
Slide 44: Expression of mAbX in different cell lines and different pHs
Expression of mAbX in 2 different cell lines and different pHs Production in (ΔXX) Comparison of six different pH-conditions 200 ng #7, pH 5.2 #8, pH 5.6 #9, pH 6.0 #10, pH 6.4 #11, pH 6.8 #12, pH 7.2 4 5 6 7 4 5 6 7 4 5 6 7 4 5 6 7 4 5 6 7 4 5 6 7 HC red LC green Fermentation of the mAX that 0.04 µ
Slide 45: Monoclonal Antibodies are Stable and Correctly Folded
Monoclonal Antibodies are Stable and Correctly Folded ➢ Typically stable mAbs production levels are Western Blotting between 5 – 10 g/L in 5-7 days 400 ng 200 ng Days 2 3 4 5 6 ➢ Best productivity at Amber system: o 24.5 g/L in 7 days (3.5 g/l/day) ➢ Best productivity so far at 30L scale: o 20.6 g/L
Slide 46: Neutralizing activity of mAb produced in fungus or CHO cells
Neutralizing activity of mAb produced in fungus or CHO cells Hemolytic assay in rabbit RBC with alpha toxin of Staphylococcus aureus Graph#1 0.2 Hemolytic assay The neutralizing activity assay 0.18 demonstrated great similarity between 0.16 C1 produced mAb and CHO produced 0.14 mAb 0.12 0.1 0.08 0.0
Slide 47: Global Health Access, Affordability & Equity
GLOBAL HEALTH ACCESS, AFFORDABILITY & EQUITY Asia North America India South America Africa Australia (including Middle East) DYADIC INFORMATION 47