A5064846835- Protein purification and stability
- Monoclonal and Polyclonal Antibodies Research
- Viral Infectious Diseases and Gene Expression in Insects
- Analytical Chemistry and Chromatography
- SARS-CoV-2 and COVID-19 Research
- Glycosylation and Glycoproteins Research
- Rare-earth and actinide compounds
- Enzyme Production and Characterization
- Transgenic Plants and Applications
- Protein Structure and Dynamics
- Bacteriophages and microbial interactions
- SARS-CoV-2 detection and testing
- Advanced Condensed Matter Physics
- Physics of Superconductivity and Magnetism
- Peptidase Inhibition and Analysis
- Microfluidic and Capillary Electrophoresis Applications
- Biochemical and Structural Characterization
- EEG and Brain-Computer Interfaces
- Biosensors and Analytical Detection
- Enzyme Structure and Function
- Machine Learning in Bioinformatics
- Ultrasonics and Acoustic Wave Propagation
- Phonocardiography and Auscultation Techniques
- Non-Destructive Testing Techniques
- HER2/EGFR in Cancer Research
Austrian Centre of Industrial Biotechnology (Austria)
2017-2024
BOKU University
2014-2024
Bioprocessing Technology Institute
2012-2013
Agency for Science, Technology and Research
2012-2013
University of Augsburg
1996-2001
Augsburg University
1999
University of Florida
1997
Freie Universität Berlin
1994
We identified active isoforms of the chimeric anti-GD2 antibody, ch14.18, a recombinant antibody produced in Chinese hamster ovary cells, which is already used clinical trials.Citation1,2,3 separated by high resolution ion-exchange chromatography with linear pH gradient elution into acidic, main and basic charge variants on preparative scale yielding enough material for an in-depth study sources effects microheterogeneity. The binding affinity toward antigen various cell surface receptors...
Fusion protein technologies to facilitate soluble expression, detection, or subsequent affinity purification in
Chimeric virus-like particles formed by the co-expression of HIV-1 gag capsid protein and surface proteins from other viruses can be used as a platform for production wide range viral vaccines. These rapidly produced in an insect cell culture using baculovirus expression vector system. However, variety different process-related impurities such host proteins, double stranded DNA, chromatin, structurally similar bionanoparticles like extracellular vesicles are present harvested supernatant...
Abstract We successfully transferred a two‐stage batch precipitation‐based antibody capture step to continuous mode using tubular reactors. The precipitation process solely employs cheap mineral salt (CaCl 2 ) and an organic solvent (ethanol) could replace the costly protein A in purification of recombinant antibodies from cell culture supernatant. time startup untill attaining steady state conditions was reached less than 15 minutes both reactors were operated for several hours at without...
It has previously been shown for individual antibodies, that the microheterogenity pattern can have a significant impact on various key characteristics of product. The aim this study to get more generalized understanding importance microheterogeneity. For purpose, charge variant different commercially available therapeutic mAb products was compared using Cation-Exchange Chromatography with linear pH gradient antigen affinity, Fc-receptor antibody dependent cellular cytotoxicity (ADCC) and...
Antibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection or vaccination. While first-generation antibody have primarily provided qualitative results, accurate seroprevalence studies and tracking of levels over time require highly specific, sensitive quantitative test setups.
This study presents a comprehensive investigation of the mechanistic understanding retention and selectivity in hydrophobic interaction chromatography. It provides valuable insights into crucial method-development parameters involved achieving chromatographic resolution for profiling molecular variants trastuzumab. Retention characteristics have been assessed three column chemistries, i.e., butyl, alkylamide, long-stranded multialkylamide ligands, while distinguishing hydrophobicity surface...
Caspase-2 is the most specific protease of all caspases and therefore highly suitable as tag removal enzyme creating an authentic N-terminus overexpressed tagged proteins interest. The wild type human caspase-2 a dimer heterodimers generated by autocatalytic processing which required for its enzymatic activity. We designed circularly permuted (cpCasp2) to overcome drawback complex recombinant expression, purification activation, cpCasp2 was constitutively active expressed single chain...
Recombinant monoclonal antibodies are predominantly produced in mammalian cell culture bioprocesses. Post-translational modifications affect the micro-heterogeneity of product and thereby influence important quality attributes, such as stability, solubility, pharmacodynamics pharmacokinetics. The analysis surface charge distribution provides aggregated information about these modifications. In this work, we established a direct injection pH gradient cation exchange chromatography method,...
Fusion protein technologies improve the expression and purification of recombinant proteins, but removal tags involved requires specific proteases. The circularly permuted caspase-2 (cpCasp2) with its cleavage site, efficiently generates untagged protein. While cpCasp2 is possible before all 20 proteinogenic amino acids, valine, leucine, isoleucine, aspartate glutamate suffers from slow, proline extremely turnover. To make platform fusion process even more general such that any an authentic...
The aim of this study was the development a scalable production process for high titer (108 pfu/mL and above) recombinant baculovirus stocks with low cell line-derived impurities virus-like particles (VLP). To achieve this, we developed density (HCD) culture footprint proliferation, compared different infection strategies at multiplicity (MOI) 0.05 0.005, validated generally applicable harvest criteria viability ≤ 80%. We also investigated online measurable parameters to observe production....
Secretory immunoglobulin A [sIgA] is a promising candidate for enteric therapeutics applications, and several sIgA-based constructs are currently being developed by groups utilizing clarified Chinese hamster ovary [CHO] cell culture supernatants. To the monoclonal antibody downstream processing typically entails chromatography-based purification processes beginning with Protein chromatography. In this paper, aqueous two-phase systems [ATPS] were employed preliminary of secretory [mAb] from...
Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and vivo because the susceptibility antibodies undergo chemical modifications. Modifications adversely affect potency drug, induce immunogenicity or pharmacokinetics. In this study, we evaluated application optimized pH gradient systems for separation variants trastuzumab after forced degradation study. gradient-based elution resulted...
Different degrees of protein purity have been observed in immobilized metal affinity chromatography ranging from extremely high to moderate and low purity. It has hypothesized that the host cell composition ligands are factors governing a obtained after (IMAC). Ni nitrilotriacetic acid (NTA) become first choice for facile His-tagged purification, but alternative such as iminodiacetic (IDA) with other ions Zn, Cu Co valuable options when expected or binding capacity is not reached. Especially...
Pertuzumab is a monoclonal antibody used for the treatment of HER2-positive breast cancer in combination with trastuzumab. Charge variants trastuzumab have been extensively described literature; however, little known about charge heterogeneity pertuzumab. Here, changes ion-exchange profile pertuzumab were evaluated by pH gradient cation-exchange chromatography after stressing it up to 3 weeks at physiological and elevated 37 °C. Isolated arising under stress conditions characterized peptide...
Secretory immunoglobulin A (sIgA) has a big potential for passive immunization and other therapeutic applications. This complex molecule with molecular mass of 410 kDa consists two IgA monomers linked by connecting chain (J) secretory component (SC). Fully assembled sIgA was overexpressed in CHO cells. In order to exploit the recombinant expressed cells platform process capture antibody directly from clarified culture supernatant using available affinity chromatography material investigated....
Biosimilars are increasing in economic importance. Just how similar a biosimilar needs to be gain market approval is currently still decided on per case basis. The authors try shed light one often cited critical quality attribute of monoclonal antibodies, namely charge heterogeneity. Using high resolution electrophoretic and chromatographic methods, the able separate quantify variant content infliximab originator three biosimilars. Additionally quantified compared antigen binding affinity an...
The Caspase-based fusion protein technology (CASPON) allows for universal cleavage of tags from proteins interest to reconstitute the native N-terminus. While CASPON enzyme has been optimized be promiscuous against a diversity N-terminal peptides, efficacy larger can surprisingly low. We develop an efficient means rationalize and predict efficiency based on structural representation intrinsically disordered peptides their putative interactions with enzyme. number favorably interacting...
Hydrophobic interaction chromatography is a versatile method to polish antibodies. Here, we present polishing procedure in order obtain an ultra-pure preparation of antitumor necrosis factor (TNF) alpha IgG1. (HIC) was used with Toyopearl® Phenyl 650 M adsorbent the presence ammonium sulfate. Adsorption isotherms, breakthrough curves and chromatographic runs were carried out. The eluted antibody recovered 99.9 % purity 96.2 yield. In main peak, aggregates, host cell proteins (HCP) DNA...