GlycoDelete - Universiteit Gent
Transcript of GlycoDelete - Universiteit Gent
GlycoDelete
Part of the Protein Expression Technology Toolbox by VIB
Koen Tyberghein, PhD
BD manager, VIB
Contact: [email protected]
BPI 2020
Confidential
Towards next-gen biopharmaceuticalsProprietary eukaryotic cell-based glyco-engineering toolbox for next-generation biopharmaceuticals
o Biobetters
o Improved pharmaceuticals (e.g. ADC’s)
Enabling tools for structural & functional analysis of challenging target proteins
Broad IP portfolio & extensive know-howo Protein glycosylation engineering: 8 patent families
o Protein expression: 2 patent families
Expanding strategic partnership base
Confidential
Heterogeneous N-glycosylation
• N-glycosylation on therapeutic proteins
o Protein purification issues
o Process reproducibility problems
o Variable therapeutic effect
o Disadvantages of engineered proteins or enzymatic approaches
Confidential
Glyco-engineering platform
Next-generation biopharmaceuticals
Proprietary glyco-engineering & proteinexpression technologies
Purification methods
Protein expression toolbox• Soluble proteins vs.
membrane proteins• Expression yield• Temporal/spatial expression
Protein analytics
EfficacyConvenienceCost
Next-gen biotherapeutics
Innovativevaccines
Glyco-engineering toolbox• Simple human N-glycans• No mucin-type O-
glycosylation
Glyco-expression systems• Mammalian (HEK293, CHO)• Yeast (Pichia pastoris)• Plants
Site-specific conjugationtechnologies• Coupling of functional
moieties (toxins, half-life extension, …)
GlycoDelete: the basic concept
Glycosylation in WT eukaryotic cells
Glycosylation in GlycoDelete cells
Confidential
Glycoengineering N-glycans O-glycans N-glycan effect
Wild type HEK293s
• heterogenic N- and O-glycans
• not function/PK optimized
GlycoDelete• well defined short N-
glycans• heterogeneous O-glycans
GlycoDoubleDelete• 1 possible N-glycan
structure• no O-glycans
GlycoDoubleDelete+Galactose
• LacNAc N-glycans for site specific coupling
• no O-glycans
Mammalian cell expression systems (HEK293, CHO)
Protein Class #
mAb 15
Viral proteins 11
Enzymes 6
Cytokines 6
Membrane proteins 6
Nanobodies 5
Antigen 1
Fc fusion 1
Total 51
Pichia technologies
Glycoengineering N-glycans N-glycan effect
Wild type Pichia pastoris
• high mannose structures, • rapid clearance• potentially immunogenic
GlycoSwitch• contaminating high mannose
structures • potentially immunogenic
GlycoSwitch followed by clean-up
• no contaminating structures • no influence on cell growth
GlycoDelete (+ GalT)
• very homogeneous glycans • starting point for chemical coupling
P
P
P
P
(+ )
Optimized biologics by glyco-engineeringGlycoDelete hIgG1
Homogeneity PharmacokineticsADCC
Fc RI receptor affinity Fc RIIa receptor affinity Fc RIIb receptor affinity
Nature Biotechnology 32, 485–489 (2014)
• Single domain antibodies as case study
• Targeted introduction of N-linked glycans in VHH scaffold
Glyco-conjugation strategiesWO/2018/206734
Glycan-specific (site selective) conjugation
GBP-R86N(1 N-glyc. site)
Periodate-based
Glyco-conjugation strategy
Galactose-oxidase based Glyco-conjugation
strategy
Enabling tools for structural & functional analysis of challenging target proteins
• Enhanced membrane protein expression
ACS Synth. Biol. 5, 1070-1075 (2016)
21 June 2019Enabling tools for structural & functional analysis of challenging target proteins
•WO 2013/107905:
“A host cell comprising a first exogenous nucleic acid sequence encoding amembrane protein and a second exogenous nucleic acid encoding ananobody that specifically binds to an extracellular or an intracellularconformational epitope of said membrane protein, each under the controlof a promoter.“
Enabling tools for structural & functional analysis of challenging target proteins
• Crystal structure of the hCSF-1:CSF-1RD1–D3 ternary complex
Structure 23, 1–11 (2015)
GlycoDelete basic patentWO2010/015722
IP strategy
Antibodies in GDWO 2015/032899
GlycoDoubleDeleteWO2017/005925
Membrane proteins in GDWO2013/107905
GlycoSwitch + endoT cleanupWO 2017/191208
Glyco-engineered Ig domains WO/2018/206734
High LacNAc in GDWO 2019/053145
High SiaLacNAc in GDWO 2019/053147
PLATFORM DEVELOPMENTS & TECHNOLOGY BUILDING
PRODUCT DEVELOPMENTS
Glyco-engineered vaccinsIn preparation
Confidential
2009 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019
Confidential
SUMMARY: Protein glycosylation engineering, technology validation
GlycoDelete in mammalian cellsNature Biotechnology 32, 485–489 (2014)GlycoDelete in plant cellsNature Biotechnology 33, 1135-7 (2015)GlycoDelete in Pichia pastorisIn preparation
Structure determination of target - Structure 23, 1–11, (2015)Production of membrane receptor - ACS Synth. Biol. 5, 1070-1075 (2016)
Production and glycan profiling of biopharmaceuticals (industry) – independently validated by 3 pharma companies, of which two now hold a non-exclusive license. One food tech company holds a non-excl. license as well.
Structure determination of challenging target proteins – validated by # structural biology labs
Technology
High impact papers
Externalvalidation
Strong IPBroad patent protection on glyco-engineered cells, and on products and applications derived thereof.
8 patent families, 3 granted patents including in US & EP
• Monoclonal Abs• Ab fragments• Membrane receptors
Different proteinclasses
• Growth factors• Enzymes• Cytokines• Hormones
Partnership opportunitiesPlatform driven or target-based
Strategic co-development partnerships
• Novel therapeutic modalities(ADCs, …)
• Improved biotherapeutics
• Bioactivity, bioavailability, immune effector functions, selectivity, …
• Target antibody discovery
• Other
License deals
• Cell line development
• mammalian, Pichia, plants
• manufacturing of biopharmaceuticals
• Enabling tools for structural& functional analysis of challenging target proteins
• Other
Contact: E - [email protected] - +32 498 63 77 86