and process optimization of biopharmaceuticals BioGlobaX Inc Anelia Atanassova Sr Scientist BioGlobaX Inc Toronto Canada August 1012 2015 London UK Drug Discovery The Traditional ID: 802887
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Analytical methodologyand toolkit for improvement of quality attributes and process optimization of biopharmaceuticals
BioGlobaX
Inc.
Anelia
Atanassova
, Sr. Scientist,
BioGlobaX
Inc., Toronto, Canada
August 10-12, 2015, London, UK
Slide2Drug DiscoveryThe Traditional: Single Drug - TargetThe “New” Biology:
X-
omics
BioGlobaX
Adopted from J. of Internal Medicine 2010, 268 pp 432-448
2
Slide3GSH/GSSHThioredoxin20 possible C-C configurations
The Human
Redox / Cys Proteome
214,000
Cys
residues
5–12% - oxidized
H. proteins 3Ds C-C bonds
Total 4,104 16,538
BioGlobaX
The Traditional:
Thiol
-based
Redox
Homeostasis
The “New” Biology:
Cys
Proteomics
3
Cook K & Hogg P (2013)
Antioxid
Redox
Signal 18: 1987–2015
Slide4The Traditional:Reactive Oxygen Species Metal Induced Oxidation and Degradation
Fe
3+
/Fe
2+
Cu
2+
/Cu
+
The “New” Biology:
Cys
Proteomics
V/S
Zn
2+
BioGlobaX
4
Slide5Antibodies:
Real Data
BioGlobaX
PDB 1HZH
PDB 4BYH
IgG1b12
IgG1
Fc
IgG1 /
Fcγ
PDB 3V7M
Sibéril
S
,
Immunol.Lett
. (2012) 143: 60-69
Zn
2+
Zn
2+
Zn
2+
5
Slide6The Traditional Approach:
C
H
3
C
H
2
C
H
1
V
H
V
L
C
L
The “New” Biology:
Redox
Proteomics
IgG
2
IgG
4
Antibodies
BioGlobaX
6
Slide7BioGlobaX7
Antibodies Abnormalities
Slide8Lessons from Enzymes: Adenylate KinaseA Paradigm ShiftsBioGlobaX8
Mg
2+
- ATP + AMP ↔ Mg
2+
- ADP + ADP
Slide9The Power of BioinformaticsBioGlobaX9
Human
AK1-trimerAP5 - Zn2+ PDB 1Z83
Slide10Analytical ToolkitBioGlobaXMethodologyBioinformaticsTargets and systemsFactors and instrumentSingle, multiple and routine assays
Sequence
Interactions 10
Slide11Analytical ToolkitBioGlobaX
Methodology
Bioinformatics
Targets and systems
Factors and instrument
Single, multiple and routine assays
Sequence
Interactions
Data Mining, Analytical & Quality
11
Slide12Factors to Consider:Factors Process ProductCell type / strains √ √Gene constructs √ √pH, T0, metabolites √ √
O2
, agitation √ √Media composition - √Sequence and structure √ √
Activity
Stability
Yield
BioGlobaX
Data Mining, Analytical & Quality
12
Slide13Factors to Consider:Factors Process ProductBuffers composition - √Purification process √ √Additives - √pH variations √ √O
2, H2
O2, ROS √ √Impurities (metals) √ √
BioGlobaX
Activity and Stability √ √
Data Mining, Analytical & Quality
13
Slide14Sequential VS DoE ApproachBioGlobaXStudy ≥ two factors in parallel; Searching for correlations between them;More precise information;
Fewer experiments.
14
Slide15Cost Effective AssaysAbsorption spectra of purified proteinsBioGlobaX15
Slide16Cost Effective AssaysAbsorption spectra of purified proteinsBioGlobaXKinetics Molecular Weight
Oligomers
/ AggregatesConformational ChangesBinding affinity / SpecificitySmall Molecule Coordination
16
Slide17Cost Effective AssaysThe Power of UV/Vis spectrometryBioGlobaX
17
Kinetics Molecular Weight
Oligomers
/ Aggregates
Conformational Changes
Binding affinity / Specificity
Small Molecule Coordination
Metal content
Oxidation status
Slide18Multidisciplinary DoE approachesRelevant data collection / bioinformatics ↔ comparative analysis
The Future
BioGlobaX
18
Slide19AcknowledgmentsBioGlobaX Inc., Canada:Slav Dimov, M.Sc.University of Toronto, Canada: Prof. Deborah Zamble Prof. Cheryl ArrowsmithProf. Aled
EdwardsProf. Emil
PaiDr. Robert LamProf. Hee-Won ParkStockholm University, Sweden:Prof.
Martin
Högbom
Nagoya University, Japan:
Prof. Masahiro
Sugiura
Prof. Mamoru Sugita
Prof.
Toshimitsu
Niwa
Sofia University, Bulgaria:
Prof.
Atanass
Atanassov
Inst. Mol. Biol., Bulgaria:
Prof. Ivan
Ivanov
Dr.
Roumjana
Mironova
Dr. Tamara
Paipanova
Dr.
Genoveva
Nacheva
www.bioglobax.com