antonio del casale microbion srl verona university...
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Microbion srl Verona University Spin-off - Italy
Antonio Del Casale
Food Matters Live – 19 November 2014
Microbion – Verona University Spin-off
Biodiversity services & products
Finds the best performing microbes using innovative DNA-fingerprinting
Microbion – Verona University Spin-off
Established company in 2011 >200 peer-review research publications
>50 Successful industrial case-studies >20 Customers >6 Industrial sectors
Technology Transfer
Microbion – Verona University Spin-off
Service provider: •Analytical services •R&D services •Bio-bank service
Products developer: •Molecular diagnostics •Pro-tech microbial strains
What we do?
Microbion – Verona University Spin-off
Innovation: •Product development •Process optimization
Quality control: •Safety assessments •Regulatory compliance
What customers need?
Microbes Biodiversity
Biodiversity Who is the good? Can the good’s win?
Microbes: Smallest & most abundant living organisms
Pathogenic Harmful
Spoiling Unwanted Anti-pro
Pro-technologic Make products Health improver
Microbes Biodiversity
Pro-technologic Make products Health improver
Natural occurring microbes
Green Chemistry • Bio-fuels • Bio-plastics • Bio-preservatives
Food • Fermented
Health • Probiotics
Agriculture • Bio-fertilisers • Bio-pesticides
Industrial microbes - Manufactured - Controlled
Microbes Biodiversity
Food Microbes
Industrial microbes • Bacteria (Lactic Acid Bacteria) • Yeast (Saccharomyces spp.)
Benefits: • Shelf-life • Taste/Flavour • Digestibility • Nutrition
+Raw ingredients
Health Microbes
Benefit: • Competition to pathogens • Wellness & Nutrition • Immuno-regulator • Psychotropics
Industrial microbes • Bacteria (Lactic Acid Bacteria) • Yeast (Saccharomyces spp.)
Human Microbiome Top 10 emerging technologies The World Economic Forum (1 September 2014)
Probiotics
Health Microbes
DNA patters Molecular markers Signature sequences Genomes sequence Meta-genomic
Advantages of the molecular approach
High discrimination High accuracy High reliability ...and OVER the species level
Intra-specie biodiversity: Same specie Different abilities
Advantages of the molecular approach
DNA patters Molecular markers Signature sequences Genomes sequence Meta-genomic
High discrimination High accuracy High reliability ...and OVER the species level
Specie
Ge
nre
STRAIN
Saccharomyces cerevisiae (strain?)
Advantages of the molecular approach
DNA patters Molecular markers Signature sequences Genomes sequence Meta-genomic
•Improved quality wines •Cost efficiency
Local population study (1000+ yeasts)
Experimental wine production (10+ yeasts)
Scale-up winemaking (2 yeasts)
MASI Yeast Bio-Bank (100+ yeasts)
Distinctive flavour
Cost saving (and C02)
Seasonal consistency
Unique on the market!
Case study: Innovation in traditional winemaking
Case study: Microbes manufacturing
• Improved quality control • Regulatory compliance • Patent deposit
Molecular Fingerprinting Services
Intra-specie biodiversity study
Manufacturing: Batch validation
Regulatory: Genome stability reference
Legal: IP right defensibility
Species
Gen
ere
Strain
?
Consumer confusion by intra-species diversity:
•S. cerevisiae pathogenic, when adapted to human niche (temperature)
•Enterococcus faecium health promoter or marker for water pollution?
•Feed additive Bacillus cereus Vs. hemotoxic B. cereus
•Probiotic Escherichia coli (Nissle 1917) Vs. virulent E. coli
genus
species
strain
Advantages of the molecular approach
Random Amplified Polymorphic DNA (RAPD-PCR)
Pulsed Field Gel Electrophoresis (PFGE)
Random genome sampling Whole genome profiling
• Rapid • Not unambiguous
• Unambiguous • Time consuming
Genome analysis
Recent advancement of PFGE methodology: Optical Mapping
Linearized genome
Genome restriction
Fragments detection and measuring
Strain specific “DNA-barcode” • Strain tracking • Detection of genomic
rearrangement Genome sequencing scaffolding
Genome analysis
L. brevis ATCC 367 (NC 008497.1) (in-silico WGM)
L. brevis KB290 (NC 020819.1) (in-silico WGM)
L. brevis ATCC 367 (NC 008497.1) (in-silico WGM)
L. brevis LMG 11437 (4 independent replicates)
Genome analysis
•Strain specific “DNA-barcode” •Genome sequencing scaffolding
Genomic DNA restriction fragment (PFGE, ribotyping)
Profiling PCR-amplified DNA (RAPD-, ERIC-, REP-PCR)
PCR-amplification of restriction fragments (AFLP)
Restriction analysis of DNA fragments (PCR-RFLP)
DNA Multi-Locus Sequence Typing (MLST)
House-keeping Multi-Locus Sequence Analysis (MLSA)
Amplified Ribosomal DNA Restriction Analysis (ARDRA)
rRNA gene sequencing analysis (SSU, LSU)
Genomic DNA-DNA Hybridisation (DDH)
Genomic DNA mol% G+C
Whole genome sequencing analysis
Genome analysis
• Tremendous advancement of DNA sequencing = Next Generation Sequencing
Reduced time and lower cost Improved downstream analysis
• Regulatory authorities suggest or require “state-of-the-art-technology = genome sequencing:
FDA 2012 - Guidance for Industry: Early Clinical Trials with Live Biotherapeutic Products: Chemistry, Manufacturing, and Control Information
EFSA 2012 - Guidance on the safety assessment of Enterococcus faecium in animal nutrition
Strain-specific diagnostics and tracking
Safety assessment
Genome analysis
• All molecular markers become available (also currently unknown) development of specific diagnostic methods
• Gene copy number and regulators of health claims relevant genes Adaptation to the intestinal niche Genes involved in the interactions with the host Probiotic factors (e.g. serpin) able to suppress inflammatory responses Antimicrobial compounds for pathogen inhibition
• Assessment of health risks Biogenic amine production Virulence genes Antibiotic-resistance gene
Genome analysis
(EFSA, 2012)
Comparative Genomics the study of the relationship of genome structure and function across different biological species or strains
Genome analysis
Antibiotic-resistant strains
Team
Antonio Del Casale – CEO •MSc Biotechnology
•Tech Transfer
•Open Innovation
Sandra Torriani – Scientific Adv. •Full Professor Microbiology
•Food-tech Innovation
• 150+ Peer review publications
Fabio Fracchetti – COO •PhD Biotechnology
•Molecular Microbiology
•Patents
Valter Carturo – CFO •MEc Business Administration •Chattered Accountant •Start-up Operations
Giovanna Felis – Scientific Adv. •Associate Professor Microbiology
•Lactic Acid Bacteria
•Taxonomy & Genomics
The leading microbiology innovation provider (micro-organisms making mega-impact)
Antonio Del Casale [email protected] www.microbion.it
Meet us at:
Milestones & Use of funds
Seed €10K Spin-off
University grants
First €10K customer
First tech partner
First conference invitation
Veneto Region grant
First commercial partner
“Impresa per Impresa” ConfindustriaProgramme
First EU-H2020 project
First patent
2011 2012 2013 2014
2015 2016 2017
Spin-out from University facilities
First patent licensed
Second patent
Marketing team
SME Instrument EU-H2020 project
First strain licensed
Third patent
Royalties > Sales Second patent licensed
Microbion Agro Microbion Health Microbion Food-tech Microbion Energy
Vitafoods Europe 2013
Golden standards for identification: Multi-locus sequence typing
Actinobacteridae putative proteomes
All vs All
BLASTp
Core genes of
Bifidobacteria
In-silico selection
One copy in every genome Dinucleotide frequencies, GC content and codon usage consistent with genome
18
Within a species, ribosomal subunit are not discriminant
9 Candidate
markers analysis (partial sequences) (Ventura et al. 2006)
7 Molecular markers
typing scheme
• High resolution between species
• Discriminate strains of same species
ROI Strategy
Potential acquisition paths: A – Management buy-back B – Partial asset sale C – Diagnostic brand co-development
•Lactic Acid Bacteria ID •Yeasts ID
Lumora – Cambridge Spin-off R&D of BART tech raised $2.1M by Tate&Lyle now licensed to 3M
Acquisition examples benchmark:
ROI Strategy
Potential acquisition paths:
Acquisition examples benchmark:
A – Management buy-back A – Management buy-back B – Partial asset sale
•Diagnostics patents •Microbial Strains
BioFire sold FilmArray asset
to bioMérieux’s for $450M
BioGaia receive €10M/year for L. reuteri from Nestlé infant nutrition
Key Advantages
2012-2013 Service and Products
20+ Customers success cases 6 Industries
Probiotics
Consumer products
Agri-food
Dairy
Wine
Universities
2013 Sales Unique advantage:
50+ collective years of experience
Market needs & solutions
Unique microbes bio-bank
Non-patented trade secrets
Open innovation network
Business Scouting…
Institutional
Key partners
Definition of Species: Group of strains sharing high level of genomic and phenotypic similarity
• Foundations for microbial characterization
• Exchange of information among scientific community • Laws regulations and guidelines FDA – Generally Regarded As Safe (GRAS) EFSA – Qualified presumption of safety (QPS)
Genome analysis