Download BioGlobaX

BioGloba
X Inc.
Analytical methodology
and toolkit for improvement of quality attributes
and process optimization of biopharmaceuticals
Anelia Atanassova, Sr. Scientist, BioGlobaX Inc., Toronto, Canada
August 10-12, 2015, London, UK
The Traditional:
Single Drug - Target
Adopted from J. of Internal Medicine 2010, 268 pp 432-448
BioGlobaX
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The Traditional: Thiol-based
Redox Homeostasis
GSH/GSSH
The “New” Biology:
Cys Proteomics
The Human Redox / Cys Proteome
214,000 Cys residues
5–12% - oxidized
Thioredoxin
H. proteins
Total
3Ds
4,104
C-C bonds
16,538
20 possible C-C configurations
Cook K & Hogg P (2013) Antioxid Redox Signal 18: 1987–2015
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The “New” Biology:
Cys Proteomics
The Traditional:
Fe3+/Fe2+
Cu2+/Cu+
BioGlobaX
V/S
Zn2+
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IgG1 / Fcγ
PDB 3V7M
Zn2+
IgG1b12
IgG1 Fc
Zn2+
Zn2+
PDB 1HZH PDB 4BYH
BioGlobaX
Sibéril S , Immunol.Lett. (2012) 143: 60-69
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The “New” Biology:
Redox Proteomics
The Traditional Approach:
VL
IgG2
CL
VH
CH1
CH2
CH3
IgG4
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BioGlobaX
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Lessons from Enzymes: Adenylate Kinase
Mg2+ - ATP + AMP ↔ Mg2+ - ADP + ADP
BioGlobaX
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Human AK1-trimer
AP5 - Zn2+ PDB 1Z83
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Methodology
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Bioinformatics
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Targets and systems
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Factors and instrument
Single, multiple and routine assays
BioGlobaX
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Sequence
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Interactions
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Data Mining, Analytical & Quality
Methodology
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Bioinformatics
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Targets and systems
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Factors and instrument
Single, multiple and routine assays
BioGlobaX
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Sequence
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Interactions
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Data Mining, Analytical & Quality
Factors
Cell type / strains
Gene constructs
pH, T0, metabolites
O2, agitation
Media composition
Sequence and structure
BioGlobaX
Process
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Product
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Activity
Stability
Yield
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Data Mining, Analytical & Quality
Factors
BioGlobaX
Process
Product
Buffers composition
Purification process
Additives
pH variations
O2, H2O2, ROS
Impurities (metals)
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Activity and Stability
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 Study ≥ two factors
in parallel;
 Searching for
correlations
between them;
 More precise
information;
 Fewer experiments.
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Absorption spectra of purified proteins
BioGlobaX
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Absorption spectra of purified proteins
BioGlobaX
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Kinetics
Molecular Weight
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Oligomers / Aggregates
Conformational Changes
Binding affinity / Specificity
Small Molecule Coordination
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The Power of UV/Vis spectrometry
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BioGlobaX
Kinetics
Molecular Weight
Oligomers / Aggregates
Conformational Changes
Binding affinity / Specificity
Small Molecule Coordination
Metal content
Oxidation status
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Relevant data collection / bioinformatics ↔ comparative analysis
Multidisciplinary DoE approaches
BioGlobaX
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www.bioglobax.com
BioGlobaX Inc., Canada:
Slav Dimov, M.Sc.
University of Toronto, Canada:
Prof. Deborah Zamble
Prof. Cheryl Arrowsmith
Prof. Aled Edwards
Prof. Emil Pai
Dr. Robert Lam
Prof. Hee-Won Park
Stockholm 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