SCIENCE

SPECIALTIES IN BRIEF

Cell line engineering
Genome engineering – Meganuclease, TALEN
Protein engineering
Protein expression and production in E. coli, yeast, insect, mammalian (HEK, CHO, sf9, Rosetta, S. cerevisiae)
Directed molecular evolution
Phage display technology
Protein purification (LC, FPLC, Profinia, HPLC)
Protein biochemistry
Protein analysis (MS, CD, Fluorescent spectroscopy, SDS-PAGE, Western, ELISA)
Cell culture technology
Fermentation, bioreactors, Celline, HyperFlask
Antibody and Cytokine engineering

CONTENTS

Major Research and R&D Projects

Fields of Scientific Experties and Research Interests

Funding

Big Pharma partnerships and contract research

Research Grants

Collaborators

MAJOR RESEARCH AND DEVELOPMENT PROJECTS

Current Projects – Coordinator and Executive Leader

Engineering cell lines for selective expression of cell surface receptor family members using targeted genome editing – multi-step TALEN-based cell line engineering (2012-2013)

Production development of  recombinant therapeutic moAb via genome engineering in CHOMeganuclease (2012)

Antigen expressing cell line development for bioassays via genome engineering – Meganuclease (2011)

Epigenetics technology standardisation for clinical diagnostics –ChIP  process controls.

Development of monoclonal antibodies for ChIP applications (2010-2012)

Cytokine production development in mammalian cells (2010-2012)

Protein engineering, in vitro evolution, drug discovery, bionanotechnology (2002-2013)

Engineering and production development of recombinant proteins (cytokines, enzymes, gene regulators, etc) for research and therapy (CHO, HEK, INSECT, YEAST, BACTERIAL production).

Antibody engineering, epitope design, libraries, hybridoma technology, HTS screening, production in  mammalian cell culture, automated purification and multi-characterisation.

Cell line engineering for protein production and bioassay development.

Technology development for functional genomics and epigenetics studies using chromatin immunoprecipitation (ChIP).

Technology platform/assay development for biopharmaceutical discovery and development.

Development of  nanodevices for drug targeting and delivery, and gene delivery – gene therapy – composite polymeric nanoparticles.

Fabrication of novel receptor proteins for bionanodevices via protein engineering – scaffold engineering, site-directed and random mutagenesis, and phage display selection.

In vitro molecular evolution of proteins for structural-functional analysis and the development of molecular tools for industrial and therapeutic applications.

Development of bioconjugation techniques for molecular erection on nanosurfaces using protein cross-linking enzymes.

Molecular characterisation and structural-functional analysis of transglutaminases (1998-2004)

Investigation of the role of the tissue-type transglutaminase in programmed cell death (apoptosis) via identification of natural substrate proteins of the enzyme using in situ labelling with novel synthetic glutamine and lysine donor peptide substrates which penetrate cells.

Molecular analysis of the mechanism of Ca++-binding and activation of the tissue-type transglutaminase using side-directed mutagenesis-based surface charge engineering, isotope equilibrium dialysis and Ca-NMR.

Investigation of the substrate specificity of the tissue-type transglutaminase using a peptide phage display approach to determine the peptide environmental requirements for cross-linkable glutamine and lysine residues.

Catalytic mechanism, molecular characterisation and structural-functional analysis of β-glucosidases, hydroxynitrile-lyases, lipid transfer proteins (1992-1998) 

Detailed reaction kinetic analysis of the mechanism of action in mixed substrate and substrate analogue inhibitor systems.

Kinetic analysis of the pH-dependence of the reaction kinetic constants.

Radioaffinity labelling of the active site and identification of catalytic residue by RP-HPLC-based peptide mapping and peptide microsequencing.

Expression in E. coli, supersecretion from yeast, characterisation or recombinant proteins.

Analysis of active centres using site-directed mutagenesis to identify catalytic and substrate-binding amino acid residues.

Expression, purification and characterisation of mutant proteins in yeast.

Homology modelling and molecular dynamics simulation-aided analysis of the mechanism of substrate and inhibitor binding in the active site of the enzyme.

Microbial biotechnology, fermentation technology (1995-1998)

Fermentation technologies for the production of insect pathogenic thread worms.

HPLC-based detection technologies for cyclic nucleotides.

Mechanism of carbon catabolite repression in Penicillium chrysogenum. 

 

FIELDS OF SCIENTIFIC EXPERTIES AND SCIENTIFIC INTERESTS

Targeted genome editing, genome engineering: Meganucleases and TALEs, KO,  KI, point mutations, gene corrections, gene fusions, gene tagging, promoter engineering. Multi-step genome engineering for complex cell line solutions for research, bioassays, and rotein production.

Cell line development and engineering: Cell transfections, flow cytometry, cell sorting, cell cloning, cell analysis, genomic sequencing (NGS) …reporters, effectors, producers

Cell-based assays: Toxicity, proliferation, cytokine bioassays, ADCC, ADCP, CDC …

Protein engineering: protein chemistry and biochemistry, protein structure and folding, enzyme kinetics and mechanism, protein characterisation, protein purification, protein structure characterisation, protein and peptide mass spectrometry, protein interactions,  immunochemistr, protein expression in bacteria, yeast, insect, and mammalian cells, computer-based molecular modelling, molecular dynamics simulations.

Antibody engineering: Therapeutic antibody development and production, Epitope mapping…

Directed molecular evolution: Random mutagenesis, EP-PCR, StEP-PCR, DNA Shuffling, protein phage display selection systems…

Protein production: E. coli, yeast, insect, mammalian (HEK, CHO, sf9, Rosetta, S. cerevisiae)

Bionanotechnology: engineering proteins for diagnostic chip devices, nanoparticle-based targeted drug delivery

Research tools (hands-on experience):

 SDS-PAGE, 2D-PAGE, HPLC, FPLC, LC, GC, CE, SPR, Fluorescent spectroscopy, CD, NMR, ELISA, MALDI-MS, HPLC-MS/MS…

Silicon Graphics, Modeller, Quanta, Sybyl…

MOLECULAR BIOLOGY – ALL EXISTING TECHNIQUES

Cell culture, Bioreactors, Flow cytometry, Cell sorting

Chromatin Immunoprecipitation – sequencing (NGS)

Complex equipment operational knowledge :

HPLC, FPLC, Profinia, AKTA

BIACORE

BD FACS ARIA III

AGILENT Bioanalyser…

User:

NGS Roche GS Junior, TECAN Evo 150, MS Q-TRAP.


FUNDING

EC, OTKA-Hungarian National Research Foundation, Biotechnology 2000, Soros Foundation, British Council, NATO, Royal Society of Great Britain, ESF, Hungarian Academy of Sciences, Bolyai Janos RF, Baross Gábor – EU Regional Development Fund, Strategic Grants

INDUSTRIAL PARTNERSHIPS AND CONTRACT RESEARCH

Engineering cell lines for selective expression of cell surface receptors using targeted genome editing (2012-2013)

Production development of  recombinant therapeutic moAb via genome engineering in CHO (2012)

Fc-receptor expressing effector cell line development for bioassays

Antigen expressing cell line development for bioassays via genome engineering (2011)

RESEARCH GRANTS

2013-present

VKSZ_12, R&D Competitiveness and Excellence Strategic Partnership Grants  – Subproject Coordinator

Research and Technology Innovation Fund. Big Pharma Partner “Cell line development using genome engineering technologies”

Budget: ~8 Million EURO , Genomics Subproject : 400,000 EURO including Cell line development.

2010-2012

BAROSS GÁBOR PROGRAM (REG_EA_KFI_09-INNANTet)Executive Project Leader 

EU N-Great Plain Regional Development Fund. „Development of innovative antibody and cytokine production technologies”

Budget: 285,000 EURO

2010-2012

BAROSS GÁBOR PROGRAM  (EA_KFI_07-EPIGEN08) – Subproject Leader

EU North-Great Plain Regional Development Fund.

„Development of epigenetic-based laboratory kits for the diagnosis of malignant diseases”

Budget:  350,000 EURO

2003-2005  

Hungarian Scientific Research Foundation OTKA, No. 43083Participant

“Molecular mechanism in the regulation of the phagocytosis in apoptotic cells”

2003-2005

Hungarian Scientific Research Foundation OTKA, No. 44798Participant

“The third synapsis. Molecular interactions between dying cells and macrophages or dendritic cells”

2002 – 2005

Biotechnology 2001, Hungarian Ministry of Education (MKM) – Subproject Coordinator

Development of gene therapy vectors for medical applications”

2000 – 2003

Hungarian Scientific Research Foundation OTKA, No. F 032994 – Project Coordinator

“Identification of natural substrates of human tissue-type transglutaminase in apoptotic cell systems

using synthetic cell-penetrating polypeptide substrates”

1997-1999

Hungarian Scientific Research Foundation OTKA, No. 023759 – Participant

“Chemical mechanism of beta-glycosidase enzymes”

 

COLLABORATIONS  (last 10 years): (companies are in italics and underlined)

International

University of Albany, Sunny, NY, USA, NanoBioscience, College of Nanosale Science and Engineering (2013-)

Prof Susan Sharfstein, CMV promoter and CHO epigenetics, http://cnse.albany.edu/AboutUs/FacultyStaff/Faculty/SusanSharfstein.aspx

Cancer Research UK, Cambridge Research Institute (2012-)

Dr Jason S. Carroll, Principal Investigator, chromatin IP tech, breast cancer diagnostics, http://www.carroll-lab.org.uk/members

Laboratory of Bioseparation Sciences, Insbruck, Austria. (2010-)

Prof András Guttman, Antibody glycosylation analysis, tech development http://www.hlbs.org/about_pi.php

Biosystems International Ltd. (2010-2012)

Prof István Kurucz, ex CSO, monoclonal antibody development, http://www.biosys-intl.com/?lg=en

University of Newcastle upon Tyne, UK, Institute for Cell and Molecular Biosciences ICaMB (1993-2006)

Prof Monica A. Hughes, ex Head of School and Director of Institute, retired, molecular genetics

Prof. Jeremy H. Lakey, Protein engineering, structural biochemistry, http://www.ncl.ac.uk/camb/staff/profile/jeremy.lakey

Prof Robert N. Lightowlers, Director of Institute, BIACORE, http://www.ncl.ac.uk/camb/staff/profile/robert.lightowlers

Dr. Joe Gray, Molecular Biology Unit, mass spectrometry, http://www.ncl.ac.uk/camb/facilities/mbu.htm

ORLA Protein Technologies Ltd. (2002-2005)

Prof. Jeremy H. Lakey, CSO, bionanosensor engineering, http://www.orlaproteins.com

Cambridge Center for Protein Engineering, MRC Center, Cambridge, UK (2002)

Sir Prof Alan R. Fersht, FRS, directed molecular evolution, http://www.mrc.ac.uk/Ourresearch/Unitscentresinstitutes/UnitCentreDetails/MRC002122

ELIZANOR Polymer, LLC (2006-2008)

Prof John Hartmann, CEO, polymeric nanoparticle-based drug delivery, http://www.elizanor.com/

Rutgers, The State University of New Jersey, Department of Pharmaceutics (2006-2008)

Prof Tamara Minko, Chair, targeted drug delivery, http://ceutics.rutgers.edu/minkohome.shtml

University of Nijmegen, Holland, Department of Biochemistry (2000)

Prof. Dr. Wilfried W. de Jong,  protein cross-linking enzymes and their substrates, retired

University of Ghent, Belgium, Department of Biochemistry, Physiology, and Microbiology (2000)

Prof. Jozef Van Beeumen, proteomics. http://www.lprobe.ugent.be/index.html

Hungary

Gedeon Richter Pharmaceuticals, Plc. Budapest, Hungary (2010-)

Dr. Zoltán Urbányi, Deputy Head of Bio-Analytical Department, cell line engineering, http://www.richter.hu

Institute of Enzymology, Hungarian Academy of Sciences, Budapest, Hungary (2013-)

Prof Beáta Vértessy, MHASc, ZFN genome engineering, http://www.enzim.hu/index.php?option=com_content&task=blogcategory&id=52&Itemid=190

Institute of Clinical Microbiology, Albert Szentgyörgyi Clinical Center, University of Szeged, Fac.Medicine (2013-)

Dr. Dezső Virók, vaccinomics, phage display, http://www.klinikaikozpont.u-szeged.hu/clinmicro/en/staff/staff/aniko-kuris.html

Internal

Medical and Health Science Center, University of Debrecen, Hungary

Department of Biochemistry and Molecular Biology (1998-)

Prof  Dr. László Fésüs, MHASc, TG2 mechanism and molecular biology, http://bmbi.med.unideb.hu/joomla15/index.php/en/staff/head-of-department

Prof Dr László Nagy, MHASc, antibody development, ChIP process development, http://nlab.med.unideb.hu/ln.htm

Research Center for Clinical Chemistry, Hungarian Academy of Sciences (2008-2009)

Prof Dr. László Muszbek, MHASc, protein cross-linking, http://crc.med.unideb.hu/lm.html

Department of Pathology (2012-)

Prof Dr Gábor Méhes, soluble receptor-based diagnostics development, http://pathol.med.unideb.hu/en/node/59

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