A5006031220- Viral Infectious Diseases and Gene Expression in Insects
- Monoclonal and Polyclonal Antibodies Research
- Protein purification and stability
- Microtubule and mitosis dynamics
- Glycosylation and Glycoproteins Research
- CAR-T cell therapy research
- Virus-based gene therapy research
- Immune cells in cancer
- Photosynthetic Processes and Mechanisms
- Immunotherapy and Immune Responses
- Biosimilars and Bioanalytical Methods
- Viral gastroenteritis research and epidemiology
- Radiopharmaceutical Chemistry and Applications
- Clinical practice guidelines implementation
- Chromosomal and Genetic Variations
- Cellular transport and secretion
- NF-κB Signaling Pathways
- Cancer Immunotherapy and Biomarkers
- Delphi Technique in Research
- Microfluidic and Bio-sensing Technologies
- Mesenchymal stem cell research
- Electrospun Nanofibers in Biomedical Applications
- Ubiquitin and proteasome pathways
- Endoplasmic Reticulum Stress and Disease
- Innovative Microfluidic and Catalytic Techniques Innovation
Boehringer Ingelheim (Germany)
2017-2023
Roche Pharma AG (Germany)
2013-2019
John Wiley & Sons (United States)
2015
Bioengineering Center
2015
National Center for Genetic Engineering and Biotechnology
2015
CS Diagnostics
2012-2013
Max Planck Society
2005-2006
Max Planck Institute of Biochemistry
2001-2006
The OLE pathway of yeast regulates the level ER-bound enzyme Delta9-fatty acid desaturase OLE1, thereby controlling membrane fluidity. A central component this regulon is transcription factor SPT23, a homolog mammalian NF-kappaB. SPT23 synthesized as an inactive, ER membrane-anchored precursor that activated by regulated ubiquitin/proteasome-dependent processing (RUP). We now show dimerizes prior to and processed molecule, p90, retains its ubiquitin modification initially remains tethered...
We developed cergutuzumab amunaleukin (CEA-IL2v, RG7813), a novel monomeric CEA-targeted immunocytokine, that comprises single IL-2 variant (IL2v) moiety with abolished CD25 binding, fused to the C-terminus of high affinity, bivalent carcinoembryonic antigen (CEA)-specific antibody devoid Fc-mediated effector functions. Its molecular design aims (i) avoid preferential activation regulatory T-cells vs. immune cells by removing binding; (ii) increase therapeutic index therapy (a) retention at...
ABSTRACT Mesenchymal stromal cells (MSCs) are promising candidates for cell therapy. Their therapeutic use requires extensive expansion to obtain a sufficiently high number of clinical applications. State‐of‐the‐art systems, that is, primarily culture flask‐based limited regarding scale‐up, automation, and reproducibility. To overcome this bottleneck, microcarrier (MC)‐based processes have been developed. For the first time, MSCs from perinatal sources umbilical cord (UC) amniotic membrane...
Abstract Process intensification strategies are needed in the field of therapeutic protein production for higher productivities, lower cost goods and improved facility utilization. This work describes an approach, which connects a tangential-flow-filtration (TFF) based pre-stage perfusion process with concentrated fed-batch culture inoculated ultra-high seeding density (uHSD). strategy shifted biomass towards pre-stage, reaching up to 45 × 10 6 cells/mL mode. Subsequently, intensified...
In recent years, coherent with growing biologics portfolios also the number of complex and thus difficult-to-express (DTE) therapeutic proteins has increased considerably. DTE challenge bioprocess development can include various protein formats such as monoclonal antibodies (mAbs), multi-specific affinity scaffolds (e.g., bispecific antibodies), cytokines, or fusion proteins. Hence, availability robust versatile Chinese hamster ovary (CHO) host cell factories is fundamental for high-yielding...
Abstract Although most therapeutic monoclonal antibodies (mAbs) can routinely be produced in the multigram per litre range, some mAb candidates turn out to difficult‐to‐express (DTE). In addition, class of more complex biological formats is permanently increasing and mammalian expression systems like Chinese hamster ovary (CHO) cell lines show low performance. Hence, there an urgent need identify any rate limiting processing step during cellular synthesis. Therefore, we assessed...
Sequence variants in recombinant biopharmaceuticals may have a relevant and unpredictable impact on clinical safety efficacy. Hence, their sensitive analysis is important throughout bioprocess development. The two stage analytical approach presented here provides quick multi clone comparison of candidate production cell lines as first stage, followed by an in-depth including identification quantitation aberrant sequence selected clones second stage. We show that the differential suitable...
The oncolytic vesicular stomatitis (VSV)-GP virus is a promising therapeutic against cancer. To ensure clinical efficacy, doses with high titers are required, which poses challenge for the manufacturing process. Perfusion cultivation processes cell densities have attracted great interest to improve production titer. This work aimed enhance titer of VSV-GP process suspension human embryonic kidney 293 (HEK293) cells by using perfusion tangential flow filtration (TFF) and retention. For this...
Abstract The development of biopharmaceutical production cell lines typically starts with generation heterogeneous populations cells, from which then single clones are established. Several regulatory guidelines require that clonal, and the actual demonstration clonality has been increasingly demanded by authorities over last years. Here, authors describe relative contribution flow cytometry mediated deposition cells in multiwell plates subsequent imaging to assurance a state art approach...
Genetically modified CHO cell lines are traditionally used for the production of biopharmaceuticals. However, an in-depth molecular understanding mechanism and exact position transgene integration into genome pharmaceutical manufacturing is still scarce. Next-generation sequencing (NGS) holds great promise strongly facilitating factories, as it has matured to a powerful affordable technology cellular genotype analysis. Targeted Locus Amplification (TLA) combined with NGS allows robust...
In-depth analytical characterization of biotherapeutics originating from different production batches is mandatory to ensure product safety and consistent molecule efficacy. Previously, we have shown unintended incorporation tyrosine (Tyr) leucine/isoleucine (Leu/Ile) at phenylalanine (Phe) positions in a recombinant produced monoclonal antibody (mAb) using an orthogonal MASCOT/SIEVE based approach for mass spectrometry data analysis. The misincorporation could be avoided by sufficient...
The analysis of data collected using design experiments (DoE) is the current gold standard to determine influence input parameters and their interactions on process performance product quality. In early development, knowledge bioprocess a new limited. Many need be investigated for thorough investigation. For eukaryotic cell cultures, intensified DoE (iDoE) has been proposed as efficient tool, requiring fewer bioreactor runs by introducing setpoint changes during bioprocess. We report first...
Interleukin-2 (IL-2) is a potent molecule in cancer therapy. Clinical application, however, limited due to its strong side effects during the treatment. We developed an IL-2 variant (IL-2v) immunocytokine circumvent drawbacks of current During production IL-2v Chinese hamster ovary (CHO) cells, molecules with fragmented and therefore reduced cytokine activity can be observed. To control product fragmentation different process conditions were investigated. By shifting temperature or pH after...
Abstract IL-2 therapy can lead to durable responses in cancer patients, but is associated with significant toxicity. None of the described IL-2-based immunocytokines has progressed beyond Phase II trials due various constraints their design: 1) pM affinity for IL-2Rαβγ on immune cells and pulmonary vascular endothelium compromising tumor targeting fusion two wildtype moieties antibody, together FcγR binding same cells; 2) Rapid systemic clearance short half-life high binding; 3) Preferential...
A high degree of charge heterogeneity is an unfavorable phenomenon commonly observed for therapeutic monoclonal antibodies (mAbs). Removal these impurities during manufacturing often comes at the cost impaired step yields. wide spectrum posttranslational and chemical modifications known to modify mAb charge. However, a deeper understanding underlying mechanisms triggering charged species would be beneficial control variants bioprocessing. In this study, comprehensive analytical investigation...
Abstract Introduction: IL-2 therapy can lead to durable responses in a modest proportion of cancer patients, but the treatment is associated with significant toxicity. Over last decades, various IL-2-based immunocytokines have been generated by fusing tumor-targeting antibodies. However, none these molecules progressed beyond Phase II trials and they are hampered liabilities: 1) High functional affinity (low pM) for IL-2Rabg on immune cells pulmonary vascular endothelium (Krieg et al., PNAS,...